Maize inbred PH2FRV

ABSTRACT

A novel maize variety designated PH2FRV and seed, plants and plant parts thereof are provided. Methods for producing a maize plant comprise crossing maize variety PH2FRV with another maize plant are provided. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into PH2FRV through backcross conversion and/or transformation, and to the maize seed, plant and plant part produced thereby are provided. Hybrid maize seed, plants or plant parts are produced by crossing the variety PH2FRV or a locus conversion of PH2FRV with another maize variety.

BACKGROUND

There are numerous steps in the development of any novel, desirablemaize variety. Plant breeding begins with the analysis and definition ofproblems and weaknesses of the current germplasm, the establishment ofprogram goals, and the definition of specific breeding objectives. Thenext step is selection of germplasm that possess the traits to meet theprogram goals. The breeder's goal is to combine in a single variety orhybrid, various desirable traits. For field crops, these traits mayinclude resistance to diseases and insects, resistance to heat anddrought, reducing the time to crop maturity, greater yield, alteredfatty acid profile, abiotic stress tolerance, improvements incompositional traits, and better agronomic characteristics and quality.

These product development processes, which lead to the final step ofmarketing and distribution, can take from six to twelve years from thetime the first cross is made until the finished seed is delivered to thefarmer for planting. Therefore, development of new varieties and hybridsis a time-consuming process that requires precise planning, efficientuse of resources, and a minimum of changes in direction. A continuinggoal of maize breeders is to develop stable, high yielding maizevarieties and hybrids that are agronomically sound with maximal yieldover one or more different conditions and environments.

SUMMARY

Provided is a novel maize, Zea mays L., variety, designated PH2FRV andprocesses for making PH2FRV. Seed of maize variety PH2FRV, plants ofmaize variety PH2FRV, plant parts and cells of maize variety PH2FRV, andprocesses for making a maize plant that comprise crossing maize varietyPH2FRV with another maize plant are provided. Also provided are maizeplants having all the physiological and morphological characteristics ofthe inbred maize variety PH2FRV.

Processes are provided for making a maize plant containing in itsgenetic material one or more traits introgressed into PH2FRV through oneor more of backcross conversion, genetic manipulation andtransformation, and to the maize seed, plant and plant parts producedthereby. Hybrid maize seed, plants or plant parts produced by crossingthe variety PH2FRV or a locus conversion of PH2FRV with another maizevariety are also provided.

The inbred maize plant may further comprise a cytoplasmic or nuclearfactor capable of conferring male sterility or otherwise preventingself-pollination, such as by self-incompatibility. Parts of the maizeplant of the present invention are also provided, for example, pollenobtained from an inbred plant and an ovule of the inbred plant.

Seed of the inbred maize variety PH2FRV is provided. The inbred maizeseed may be an essentially homogeneous population of inbred maize seedof the variety designated PH2FRV. Essentially homogeneous populations ofinbred seed are generally free from substantial numbers of other seed.Therefore, inbred seed generally forms at least about 97% of the totalseed. The population of inbred maize seed of the invention may beparticularly defined as being essentially free from hybrid seed. Theinbred seed population may be separately grown to provide an essentiallyhomogeneous population of inbred maize plants designated PH2FRV.

Compositions are provided comprising a seed of maize variety PH2FRVcomprised in plant seed growth media. In certain embodiments, the plantseed growth media is a soil or synthetic cultivation medium. In specificembodiments, the growth medium may be comprised in a container or may,for example, be soil in a field.

A plant of maize variety PH2FRV comprising an added heritable trait isprovided. The heritable trait may comprise a genetic locus that is adominant or recessive allele. In one embodiment, a plant of maizevariety PH2FRV comprising a single locus conversion in particular isprovided. An added genetic locus which confers one or more traits suchas, for example, male sterility, herbicide tolerance, insect resistance,disease resistance, waxy starch, modified fatty acid metabolism,modified phytic acid metabolism, modified carbohydrate metabolism andmodified protein metabolism is provided. The trait may be, for example,conferred by a naturally occurring maize gene introduced into the genomeof the variety by backcrossing, a natural or induced mutation, or atransgene introduced through genetic transformation techniques. Whenintroduced through transformation, a genetic locus may comprise one ormore transgenes integrated at a single chromosomal location.

An inbred maize plant of the variety designated PH2FRV is provided,wherein a cytoplasmically-inherited trait has been introduced into saidinbred plant. Such cytoplasmically-inherited traits are passed toprogeny through the female parent in a particular cross. An exemplarycytoplasmically-inherited trait is the male sterility trait.Cytoplasmic-male sterility (CMS) is a pollen abortion phenomenondetermined by the interaction between the genes in the cytoplasm and thenucleus. Alteration in the mitochondrial genome and the lack of restorergenes in the nucleus will lead to pollen abortion. With either a normalcytoplasm or the presence of restorer gene(s) in the nucleus, the plantwill produce pollen normally. A CMS plant can be pollinated by amaintainer version of the same variety, which has a normal cytoplasm butlacks the restorer gene(s) in the nucleus, and continue to be malesterile in the next generation. The male fertility of a CMS plant can berestored by a restorer version of the same variety, which must have therestorer gene(s) in the nucleus. With the restorer gene(s) in thenucleus, the offspring of the male-sterile plant can produce normalpollen grains and propagate. A cytoplasmically inherited trait may be anaturally occurring maize trait or a trait introduced through genetictransformation techniques.

A tissue culture of regenerable cells of a plant of variety PH2FRV isprovided. The tissue culture can be capable of regenerating plantscapable of expressing all of the physiological and morphological orphenotypic characteristics of the variety, and of regenerating plantshaving substantially the same genotype as other plants of the variety.Examples of some of the physiological and morphological characteristicsthat may be assessed include characteristics related to yield, maturity,and kernel quality. The regenerable cells in such tissue cultures can bederived, for example, from embryos, meristematic cells, immaturetassels, microspores, pollen, leaves, anthers, roots, root tips, silk,flowers, kernels, ears, cobs, husks, or stalks, or from callus orprotoplasts derived from those tissues. Maize plants regenerated fromthe tissue cultures of the invention, the plants having all thephysiological and morphological characteristics of variety PH2FRV arealso provided.

Processes are provided for producing maize seeds or plants, whichprocesses generally comprise crossing a first parent maize plant as amale or female parent with a second parent maize plant, wherein at leastone of the first or second parent maize plants is a plant of the varietydesignated PH2FRV. These processes may be further exemplified asprocesses for preparing hybrid maize seed or plants, wherein a firstinbred maize plant is crossed with a second maize plant of a different,distinct variety to provide a hybrid that has, as one of its parents,the inbred maize plant variety PH2FRV. In these processes, crossing willresult in the production of seed. The seed production occurs regardlessof whether the seed is collected or not.

In some embodiments, the first step in “crossing” comprises planting,such as in pollinating proximity, seeds of a first and second parentmaize plant, and such as, seeds of a first inbred maize plant and asecond, distinct inbred maize plant. Where the plants are not inpollinating proximity, pollination can nevertheless be accomplished bytransferring a pollen or tassel bag from one plant to the other asdescribed below.

A second step comprises cultivating or growing the seeds of said firstand second parent maize plants into plants that bear flowers (maizebears both male flowers (tassels) and female flowers (silks) in separateanatomical structures on the same plant).

A third step comprises preventing self-pollination of the plants, i.e.,preventing the silks of a plant from being fertilized by any plant ofthe same variety, including the same plant. This can be done byemasculating the male flowers of the first or second parent maize plant,(i.e., treating or manipulating the flowers so as to prevent pollenproduction, in order to produce an emasculated parent maize plant).Self-incompatibility systems may also be used in some hybrid crops forthe same purpose. Self-incompatible plants still shed viable pollen andcan pollinate plants of other varieties but are incapable of pollinatingthemselves or other plants of the same variety.

A fourth step may comprise allowing cross-pollination to occur betweenthe first and second parent maize plants. When the plants are not inpollinating proximity, this is done by placing a bag, usually paper orglassine, over the tassels of the first plant and another bag over thesilks of the incipient ear on the second plant. The bags are left inplace for at least 24 hours. Since pollen is viable for less than 24hours, this assures that the silks are not pollinated from other pollensources, that any stray pollen on the tassels of the first plant isdead, and that the only pollen transferred comes from the first plant.The pollen bag over the tassel of the first plant is then shakenvigorously to enhance release of pollen from the tassels, and the shootbag is removed from the silks of the incipient ear on the second plant.Finally, the pollen bag is removed from the tassel of the first plantand is placed over the silks of the incipient ear of the second plant,shaken again and left in place. Yet another step comprises harvestingthe seeds from at least one of the parent maize plants. The harvestedseed can be grown to produce a maize plant or hybrid maize plant.

Also provided are maize seed and plants produced by a process thatcomprises crossing a first parent maize plant with a second parent maizeplant, wherein at least one of the first or second parent maize plantsis a plant of the variety designated PH2FRV. In one embodiment of theinvention, maize seed and plants produced by the process are firstgeneration (F1) hybrid maize seed and plants produced by crossing aninbred in accordance with the invention with another, distinct inbred.Seed of an F1 hybrid maize plant is contemplated and an F1 hybrid maizeplant and seed thereof are provided.

The genetic complement of the maize plant variety designated PH2FRV isprovided. The phrase “genetic complement” is used to refer to theaggregate of nucleotide sequences, the expression of which sequencesdefines the phenotype of, in the present case, a maize plant, or a cellor tissue of that plant. A genetic complement thus represents thegenetic make-up of an inbred cell, tissue or plant, and a hybrid geneticcomplement represents the genetic make-up of a hybrid cell, tissue orplant. Maize plant cells that have a genetic complement in accordancewith the inbred maize plant cells disclosed herein, and plants, seedsand diploid plants containing such cells are provided.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that variety PH2FRV could be identified by any of themany well-known techniques used for genetic profiling disclosed herein.

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by maize plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of an inbred maize plant of the invention with a haploidgenetic complement of a second maize plant, such as, another, distinctinbred maize plant. In another aspect, the present invention provides amaize plant regenerated from a tissue culture that comprises a hybridgenetic complement of this invention.

Methods of producing an inbred maize plant derived from the maizevariety PH2FRV are provided, the method comprising the steps of: (a)preparing a progeny plant derived from maize variety PH2FRV, whereinsaid preparing comprises crossing a plant of the maize variety PH2FRVwith a second maize plant; (b) crossing the progeny plant with itself ora second plant to produce a seed of a progeny plant of a subsequentgeneration; (c) repeating steps (a) and (b) with sufficient inbreedinguntil a seed of an inbred maize plant derived from the variety PH2FRV isproduced. In the method, it may be desirable to select particular plantsresulting from step (c) for continued crossing according to steps (b)and (c). By selecting plants having one or more desirable traits, aninbred maize plant derived from the maize variety PH2FRV is obtainedwhich possesses some of the desirable traits of maize variety PH2FRV aswell as potentially other selected traits.

DETAILED DESCRIPTION

A new and distinctive maize inbred variety designated PH2FRV, which hasbeen the result of years of careful breeding and selection in acomprehensive maize breeding program is provided.

Definitions

Maize (Zea mays) can be referred to as maize or corn. Certaindefinitions used in the specification are provided below. Also in theexamples that follow, a number of terms are used herein. In order toprovide a clear and consistent understanding of the specification andclaims, including the scope to be given such terms, the followingdefinitions are provided. NOTE: ABS is in absolute terms and % MN ispercent of the mean for the experiments in which the inbred or hybridwas grown. PCT designates that the trait is calculated as a percentage.% NOT designates the percentage of plants that did not exhibit a trait.For example, STKLDG % NOT is the percentage of plants in a plot thatwere not stalk lodged. These designators will follow the descriptors todenote how the values are to be interpreted. Below are the descriptorsused in the data tables included herein.

ABIOTIC STRESS TOLERANCE: resistance to non-biological sources of stressconferred by traits such as nitrogen utilization efficiency, alterednitrogen responsiveness, drought resistance, cold, and salt resistance

ABTSTK=ARTIFICIAL BRITTLE STALK: A count of the number of “snapped”plants per plot following machine snapping. A snapped plant has itsstalk completely snapped at a node between the base of the plant and thenode above the ear. Expressed as percent of plants that did not snap.

ALLELE: Any of one or more alternative forms of a genetic sequence. In adiploid cell or organism, the two alleles of a given sequence typicallyoccupy corresponding loci on a pair of homologous chromosomes.

ALTER: The utilization of up-regulation, down-regulation, or genesilencing.

ANTHESIS: The time of a flower's opening.

ANTIOXIDANT: A chemical compound or substance that inhibits oxidation,including but not limited to tocopherol or tocotrienols.

ANT ROT=ANTHRACNOSE STALK ROT (Colletotrichum graminicola): A 1 to 9visual rating indicating the resistance to Anthracnose Stalk Rot. Ahigher score indicates a higher resistance. Data are collected only whensufficient selection pressure exists in the experiment measured.

BACKCROSSING: Process in which a breeder crosses a hybrid progenyvariety back to one of the parental genotypes one or more times.

BACKCROSS PROGENY: Progeny plants produced by crossing one maize line(recurrent parent) with plants of another maize line (donor) thatcomprise a desired trait or locus, selecting progeny plants thatcomprise the desired trait or locus, and crossing them with therecurrent parent one or more times to produce backcross progeny plantsthat comprise said trait or locus.

BARPLT=BARREN PLANTS: The percent of plants per plot that were notbarren (lack ears).

BLUP=BEST LINEAR UNBIASED PREDICTION. The BLUP values are determinedfrom a mixed model analysis of hybrid performance observations atvarious locations and replications. BLUP values for inbred maize plants,breeding values, are estimated from the same analysis using pedigreeinformation.

BORBMN=ARTIFICIAL BRITTLE STALK MEAN: The mean percent of plants not“snapped” in a plot following artificial selection pressure. A snappedplant has its stalk completely snapped at a node between the base of theplant and the node above the ear. Expressed as percent of plants thatdid not snap. A higher number indicates better tolerance to brittlesnapping.

BRENGMN=BRITTLE STALK ENERGY MEAN: The mean amount of energy per unitarea needed to artificially brittle snap a corn stalk. A high numberindicates better tolerance to brittle snapping.

BREEDING: The genetic manipulation of living organisms.

BREEDING CROSS: A cross to introduce new genetic material into a plantfor the development of a new variety. For example, one could cross plantA with plant B, wherein plant B would be genetically different fromplant A. After the breeding cross, the resulting F1 plants could then beselfed or sibbed for one, two, three or more times (F1, F2, F3, etc.)until a new inbred variety is developed.

BREEDING VALUE: A relative value determined by evaluating the progeny ofthe parent. For corn the progeny is often the F1 generation and theparent is often an inbred variety.

BRLPNE=ARTIFICIAL ROOT LODGING EARLY SEASON: The percent of plants notroot lodged in a plot following artificial selection pressure appliedprior to flowering. A plant is considered root lodged if it leans fromthe vertical axis at an approximately 30 degree angle or greater.Expressed as percent of plants that did not root lodge. A high numberindicates higher tolerance to root lodging.

BRLPNL=ARTIFICIAL ROOT LODGING LATE SEASON: The percent of plants notroot lodged in a plot following artificial selection pressure duringgrain fill. A plant is considered root lodged if it leans from thevertical axis at an approximately 30 degree angle or greater. Expressedas percent of plants that did not root lodge. A high number indicateshigher tolerance to root lodging.

BRTSTK=BRITTLE STALKS: This is a measure of the stalk breakage near thetime of pollination, and is an indication of whether a hybrid or inbredwould snap or break near the time of flowering under severe winds. Dataare presented as percentage of plants that did not snap. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

BRTPCN=BRITTLE STALKS: This is an estimate of the stalk breakage nearthe time of pollination, and is an indication of whether a hybrid orinbred would snap or break near the time of flowering under severewinds. Data are presented as percentage of plants that did not snap.Data are collected only when sufficient selection pressure exists in theexperiment measured.

CARBOHYDRATE: Organic compounds comprising carbon, oxygen and hydrogen,including sugars, starches and cellulose.

CELL: Cell as used herein includes a plant cell, whether isolated, intissue culture or incorporated in a plant or plant part.

CLDTST=COLD TEST: The percent of plants that germinate under cold testconditions.

CLN=CORN LETHAL NECROSIS: Synergistic interaction of maize chloroticmottle virus (MCMV) in combination with either maize dwarf mosaic virus(MDMV-A or MDMV-B) or wheat streak mosaic virus (WSMV). A 1 to 9 visualrating indicating the resistance to Corn Lethal Necrosis. A higher scoreindicates a higher resistance. Data are collected only when sufficientselection pressure exists in the experiment measured.

CMSMT=COMMON SMUT: This is the percentage of plants not infected withCommon Smut. Data are collected only when sufficient selection pressureexists in the experiment measured.

COMRST=COMMON RUST (Puccinia sorghi): A 1 to 9 visual rating indicatingthe resistance to Common Rust. A higher score indicates a higherresistance. Data are collected only when sufficient selection pressureexists in the experiment measured.

CROSS POLLINATION: Fertilization by the union of two gametes fromdifferent plants.

CROSSING: The combination of genetic material by traditional methodssuch as a breeding cross or backcross, but also including protoplastfusion and other molecular biology methods of combining genetic materialfrom two sources.

D and D1-Dn: represents the generation of doubled haploid.

D/D=DRYDOWN: This represents the relative rate at which a hybrid willreach acceptable harvest moisture compared to other hybrids on a 1 to 9rating scale. A high score indicates a hybrid that dries relatively fastwhile a low score indicates a hybrid that dries slowly.

DIGENG=DIGESTIBLE ENERGY: Near-infrared transmission spectroscopy, NIT,prediction of digestible energy.

DIPERS=DIPLODIA EAR MOLD SCORES (Diplodia maydis and Diplodiamacrospora): A 1 to 9 visual rating indicating the resistance toDiplodia Ear Mold. A higher score indicates a higher resistance. Dataare collected only when sufficient selection pressure exists in theexperiment measured.

DIPLOID PLANT PART: Refers to a plant part or cell that has a samediploid genotype.

DIPROT=DIPLODIA STALK ROT SCORE: Score of stalk rot severity due toDiplodia (Diplodia maydis). Expressed as a 1 to 9 score with 9 beinghighly resistant. Data are collected only when sufficient selectionpressure exists in the experiment measured.

DRPEAR=DROPPED EARS: A measure of the number of dropped ears per plotand represents the percentage of plants that did not drop ears prior toharvest. Data are collected only when sufficient selection pressureexists in the experiment measured.

D/T=DROUGHT TOLERANCE: This represents a 1 to 9 rating for droughttolerance, and is based on data obtained under stress conditions. A highscore indicates good drought tolerance and a low score indicates poordrought tolerance. Data are collected only when sufficient selectionpressure exists in the experiment measured.

EARMLD=GENERAL EAR MOLD: Visual rating (1 to 9 score) where a 1 is verysusceptible and a 9 is very resistant. This is based on overall ratingfor ear mold of mature ears without determining the specific moldorganism, and may not be predictive for a specific ear mold. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

EARHT=EDEARHT=EAR HEIGHT: The ear height is a measure from the ground tothe highest placed developed ear node attachment and is measured ininches (EARHT) or centimeters (EDEARHT).

EARSZ=EAR SIZE: A 1 to 9 visual rating of ear size. The higher therating the larger the ear size.

EBTSTK=EARLY BRITTLE STALK: A count of the number of “snapped” plantsper plot following severe winds when the corn plant is experiencing veryrapid vegetative growth in the V5-V8 stage. Expressed as percent ofplants that did not snap. Data are collected only when sufficientselection pressure exists in the experiment measured.

ECB1 LF=EUROPEAN CORN BORER FIRST GENERATION LEAF FEEDING (Ostrinianubilalis): A 1 to 9 visual rating indicating the resistance topreflowering leaf feeding by first generation European Corn Borer. Ahigher score indicates a higher resistance. Data are collected only whensufficient selection pressure exists in the experiment measured.

ECB2IT=EUROPEAN CORN BORER SECOND GENERATION INCHES OF TUNNELING(Ostrinia nubilalis): Average inches of tunneling per plant in thestalk. Data are collected only when sufficient selection pressure existsin the experiment measured.

ECB2SC=EUROPEAN CORN BORER SECOND GENERATION (Ostrinia nubilalis): A 1to 9 visual rating indicating post flowering degree of stalk breakageand other evidence of feeding by second generation European Corn Borer.A higher score indicates a higher resistance. Data are collected onlywhen sufficient selection pressure exists in the experiment measured.

ECBDPE=EUROPEAN CORN BORER DROPPED EARS (Ostrinia nubilalis): Droppedears due to European Corn Borer. Percentage of plants that did not dropears under second generation European Corn Borer infestation. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

ECBLSI=EUROPEAN CORN BORER LATE SEASON INTACT (Ostrinia nubilalis): A 1to 9 visual rating indicating late season intactness of the corn plantgiven damage (stalk breakage above and below the top ear) causedprimarily by 2^(nd) and/or 3^(rd) generation ECB larval feeding beforeharvest. A higher score indicates more intact plants. Data are collectedonly when sufficient selection pressure exists in the experimentmeasured.

EDANTCOLs=ANTHER COLOR: Rated on a 1 to 7 scale where 1 is green, 2 isyellow, 3 is pink, 5 is red, and 7 is purple.

EDantants=ANTHER ANTHOCYANIN COLOR INTENSITY: A measure of antheranthocyanin color intensity rated on a 1 to 9 scale where 1 is absent orvery weak, 3 is weak, 5 is medium, 7 is strong, and 9 is very strong.Observed in the middle third of the main branch on fresh anthers.

EDbarants=GLUME ANTHOCYANIN COLORATION AT BASE (WHOLE PLANT, EARINSERTION LEVEL): A measure of the color intensity at the base of theglume, rated on a 1 to 9 scale where 1 is absent or very weak, 3 isweak, 5 is medium, 7 is strong, and 9 is very strong. Observed in themiddle third of the main branch of the tassel.

EDBARCOLs=BAR GLUME COLOR INTENSITY: A measure of the bar glume colorintensity. Bar glume is a dark purple band that may occur on the bottomof a glume. Bar glume color intensity is measured on a scale of 1 to 7where 1 is absent, 2 is weak, 3 is medium, 5 is strong, and 7 is verystrong.

EDBRROANTs=BRACE ROOTS ANTHOCYANIN COLORATION: A measure of the colorintensity of the brace roots rated on a 1 to 9 scale where 1 is absentor very weak, 3 is weak, 5 is medium, 7 is strong, and 9 is very strong.Observed when well developed and fresh brace roots are present on 50% ofplants.

EDCOBAINTs=COB GLUME ANTHOCYANIN COLOR INTENSITY: Rated on a 1 to 9scale where 1 is absent or very weak, 3 is weak, 5 is medium, 7 isstrong, and 9 is very strong. Anthocyanin coloration should be observedon the middle third of the uppermost cob, after the removal of some ofthe grains.

EDCOBCOLs=COB COLOR: A measure of the intensity of pink or salmoncoloration of the cob, rated on a 1 to 9 scale where 1 is absent orwhite, 2 is light pink, 3 is pink, 4 is medium red, 5 is red, 6 ismedium red, 7 is dark red, 8 is dark to very dark red, and 9 is present.

EDCOBDIA=COB DIAMETER: Measured in mm.

EDCOBICAs=COB ANTHOCYANIN COLOR INTENSITY: A measure of the intensity ofpink or salmon coloration of the cob, rated on a 1 to 9 scale where 1 isvery weak, 3 is weak, 5 is medium, 7 is strong, and 9 is very strong.

EDEARDIA=EAR DIAMETER: Measured in mm.

EDEARHULs=EAR HUSK LENGTH: A measure of ear husk length rated on a 1 to9 scale where 1 is very short, 3 is short, 5 is medium, 7 is long, and 9is very long.

EDEARLNG=EAR LENGTH: Measured in mm.

EDEARROW=NUMBER OF ROWS OF GRAIN ON EAR.

EDEARSHAs=EAR SHAPE (TAPER): Rated on a 1 to 3 scale where 1 is conical,2 is conico-cylindrical, and 3 is cylindrical.

EDEARSHLs=EAR SHANK LENGTH SCALE: A measure of the length of the earshank or peduncle, rated on a 1 to 9 scale where 1 is very short, 3 isshort, 5 is medium, 7 is long, 9 is very long.

EDFILEANs=SHEATH ANTHOCYANIN COLOR INTENSITY AT FIRST LEAF STAGE: Ameasure of the anthocyanin color intensity of the sheath of the firstleaf, rated on a 1 to 9 scale where 1 is absent or very weak, 3 is weak,5 is medium, 7 is strong, and 9 is very strong.

EDFILECOs=FOLIAGE INTENSITY OF GREEN COLOR: A measure of the greencoloration intensity in the leaves, rated on a 1 to 3 scale where 1 islight, 2 is medium, and 3 is dark.

EDFILESHs=LEAF TIP SHAPE: An indication of the shape of the apex of thefirst leaf, rated on a 1 to 5 scale where 1 is pointed, 2 is pointed torounded, 3 is rounded, 4 is rounded to spatulate, and 5 is spatulate.

EDGLUANTs=GLUME ANTHOCYANIN COLOR EXCLUDING BASE: A measure of the colorintensity of the glume excluding the base, rated on a 1 to 9 scale where1 is absent or very weak, 3 is weak, 5 is medium, 7 is strong, and 9 isvery strong. Observed in the middle third of the main branch of thetassel.

EDGLUCOLs=GLUME COLOR: Rated on a 1 to 7 scale where 1 is green, 2 isyellow, 3 is pink, 5 is red, and 7 is purple.

EDKERDOCs=DORSAL SIDE OF GRAIN COLOR: Rated on a 1 to 10 scale where 1is white, 2 is yellowish white, 3 is yellow, 4 is yellow orange, 5 isorange, 6 is red orange, 7 is red, 8 is purple, 9 is brownish, and 10 isblue black. Observed in the middle third of the uppermost ear when welldeveloped.

EDKERSHAs=KERNEL SHAPE: Rated on a 1 to 3 scale where 1 is round, 2 iskidney-shaped, and 3 is cuneiform.

EDKERTCOs=TOP OF GRAIN COLOR: Rated on a 1 to 10 scale where 1 is white,2 is yellowish white, 3 is yellow, 4 is yellow orange, 5 is orange, 6 isred orange, 7 is red, 8 is purple, 9 is brownish, and 10 is blue black.Observed in the middle third of the uppermost ear when well developed.

EDLEAANGs=LEAF ANGLE BETWEEN BLADE AND STEM: A measure of the angleformed between stem and leaf, rated on a 1 to 9 scale where 1 is verysmall (<5 degrees), 3 is small (6 to 37 degrees), 5 is medium (38 to 62degrees), 7 is large (63 to 90 degrees), and 9 is very large (>90degrees). Observed on the leaf just above the upper ear.

EDLEAATTs=LEAF ATTITUDE OF ENTIRE PLANT: A measure of leaf curvature orattitude, rated on a 1 to 9 scale where 1 is absent or very slightlyrecurved, 3 is slightly recurved, 5 is moderately recurved, 7 isstrongly recurved, and 9 is very strongly recurved. Observed on the leafjust above the upper ear.

EDLEALNGs=LEAF LENGTH SCORE: A measure of leaf length rated on a 1 to 9scale where 1 indicates <0.70 m, 3 indicates 0.70 m to 0.80 m, 5indicates 0.80 m to 0.90 m, 7 indicates 0.90 m to 1 m, and 9indicates >1.00 m.

EDLEAWID=LEAF WIDTH OF BLADE: A measure of the average leaf width incentimeters.

EDLELIANTs=LEAF LIMB ANTHOCYANIN COLOR INTENSITY OF ENTIRE PLANT: Ameasure of the leaf limb anthocyanin coloration, rated on a 1 to 9 scalewith 1 being absent or very weak, 3 being weak, 5 being medium, 7 beingstrong, and 9 being very strong.

EDNODANTS=NODES ANTHOCYANIN COLOR INTENSITY: A measure of theanthocyanin coloration of nodes, rated on a 1 to 9 scale where 1 isabsent or very weak, 3 is weak, 5 is medium, 7 is strong, and 9 is verystrong.

EDRATIOEP=RATIO HEIGHT OF INSERTION OF PEDUNCLE OF UPPER EAR TO PLANTLENGTH.

EDSHEAHAs=LEAF SHEATH HAIRNESS SCALE: Rated on a 1 to 6 scale where 1indicates none and 6 indicates fuzzy.

EDSHEAANTs=SHEATH ANTHOCYANIN COLOR INTENSITY: Rated on a 1 to 9 scalewhere 1 is absent or very weak, 3 is weak, 5 is medium, 7 is strong, and9 is very strong.

EDSLKAINTs=SILK ANTHOCYANIN COLOR INTENSITY: A measure of the colorintensity of the silks, rated on a 1 to 9 scale where 1 is absent orvery weak, 3 is weak, 5 is medium, 7 is strong, and 9 is very strong.

EDSTLANTs=INTERNODE ANTHOCYANIN COLOR INTENSITY: A measure ofanthocyanin coloration of nodes, rated on a 1 to 9 scale where 1 isabsent or very weak, 3 is weak, 5 is medium, 7 is strong, and 9 is verystrong. Observed just above the insertion point of the peduncle of theupper ear.

EDTA1 RYATs=TASSEL LATERAL BRANCH CURVATURE: Rated on a 1 to 9 scalewhere 1 indicates absent or very slightly recurved (<5 degrees), 3indicates slightly recurved (6 to 37 degrees), 5 indicates moderatelyrecurved (38 to 62 degrees), 7 indicates strongly recurved (63 to 90degrees), and 9 indicates very strongly recurved (>90 degrees). Observedon the second branch from the bottom of the tassel.

EDTA1RYBRs=NUMBER OF PRIMARY LATERAL TASSEL BRANCHES: Rated on a 1 to 9scale where 1 indicates absent or very few (<4 branches), 3 indicatesfew (4 to 10), 5 indicates medium (11 to 15), 7 indicates many (16 to20), and 9 indicates very many (>20).

EDTASAHB=LENGTH OF MAIN AXIS ABOVE HIGHEST LATERAL BRANCH: The length ofthe tassel's main axis above the highest lateral branch in centimeters.

EDTASANGs=TASSEL ANGLE BETWEEN MAIN AXIS AND LATERAL BRANCHES: Rated ona 1 to 9 scale where 1 is very small (<5 degrees), 3 is small (6 to 37degrees), 5 is medium (38 to 62 degrees), 7 is large (63 to 90 degrees),and 9 is very large (>90 degrees). Observed on the second branch fromthe bottom of the tassel.

EDTASEBRs=SECONDARY TASSEL BRANCHES (NUMBER): The number of secondarytassel branches, rated on a 1 to 7 scale where 1 indicates 0 to 3branches, 2 indicates 4 to 10, 3 indicates 11 to 15, 5 indicates 16 to20, and 7 indicates >20.

EDTASLPBRs=PRIMARY TASSEL BRANCH LENGTH: A measure of the length of theprimary or lateral tassel branch, rated on a 1 to 9 scale where 1 isvery short, 3 is short, 5 is medium, 7 is long, 9 is very long. Observedon the second branch from the bottom of the tassel.

EDTASULB=LENGTH OF MAIN AXIS ABOVE LOWEST LATERAL BRANCH: The length ofthe tassel's main axis above the lowest lateral branch in centimeters.

EDZIGZAGs=DEGREE OF STEM ZIG-ZAG: Rated on a scale of 1 to 3 where 1 isabsent or very slight, 2 is slight, and 3 is strong.

EGRWTH=EARLY GROWTH: This is a measure of the relative height and sizeof a corn seedling at the 2-4 leaf stage of growth. This is a visualrating (1 to 9), with 1 being weak or slow growth, 5 being averagegrowth and 9 being strong growth. Taller plants, wider leaves, moregreen mass and darker color constitute higher score. Data are collectedonly when sufficient selection pressure exists in the experimentmeasured.

ERTLDG=EARLY ROOT LODGING: The percentage of plants that do not rootlodge prior to or around anthesis; plants that lean from the verticalaxis at an approximately 30 degree angle or greater would be counted asroot lodged. Data are collected only when sufficient selection pressureexists in the experiment measured.

ERTLPN=EARLY ROOT LODGING: An estimate of the percentage of plants thatdo not root lodge prior to or around anthesis; plants that lean from thevertical axis at an approximately 30 degree angle or greater would beconsidered as root lodged. Data are collected only when sufficientselection pressure exists in the experiment measured.

ERTLSC=EARLY ROOT LODGING SCORE: Score for severity of plants that leanfrom a vertical axis at an approximate 30 degree angle or greater whichtypically results from strong winds prior to or around floweringrecorded within 2 weeks of a wind event. Expressed as a 1 to 9 scorewith 9 being no lodging. Data are collected only when sufficientselection pressure exists in the experiment measured.

ESSENTIAL AMINO ACIDS: Amino acids that cannot be synthesized by anorganism and therefore must be supplied in the diet.

ESTCNT=EARLY STAND COUNT: This is a measure of the stand establishmentin the spring and represents the number of plants that emerge on perplot basis for the inbred or hybrid.

EXPRESSING: Having the genetic potential such that under the rightconditions, the phenotypic trait is present.

EXTSTR=EXTRACTABLE STARCH: Near-infrared transmission spectroscopy, NIT,prediction of extractable starch.

EYESPT=EYE SPOT (Kabatiella zeae or Aureobasidium zeae): A 1 to 9 visualrating indicating the resistance to Eye Spot. A higher score indicates ahigher resistance. Data are collected only when sufficient selectionpressure exists in the experiment measured.

FATTY ACID: A carboxylic acid (or organic acid), often with a longaliphatic tail (long chains), either saturated or unsaturated.

F1 PROGENY: A progeny plant produced by crossing a plant of one maizeline with a plant of another maize line.

FUSERS=FUSARIUM EAR ROT SCORE (Fusarium moniliforme or Fusariumsubglutinans): A 1 to 9 visual rating indicating the resistance toFusarium Ear Rot. A higher score indicates a higher resistance. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

GDU=GROWING DEGREE UNITS: Using the Barger Heat Unit Theory, whichassumes that maize growth occurs in the temperature range 50 degreesF.-86 degrees F. and that temperatures outside this range slow downgrowth; the maximum daily heat unit accumulation is 36 and the minimumdaily heat unit accumulation is 0. The seasonal accumulation of GDU is amajor factor in determining maturity zones.

GDUSHD=EDDAYSH=GDU TO SHED: The number of growing degree units (GDUs) orheat units required for an inbred variety or hybrid to haveapproximately 50 percent of the plants shedding pollen and is measuredfrom the time of planting. Growing degree units are calculated by theBarger Method, where the heat units for a 24-hour period are:GDU=(Max. temp.+Min. temp.)−50

The units determined by the Barger Method are then divided by 10. Thehighest maximum temperature used is 86 degrees F. and the lowest minimumtemperature used is 50 degrees F. For each inbred or hybrid it takes acertain number of GDUs to reach various stages of plant development.

GDUSLK=EDDAYSLK=GDU TO SILK: The number of growing degree units requiredfor an inbred variety or hybrid to have approximately 50 percent of theplants with silk emergence from time of planting. Growing degree unitsare calculated by the Barger Method as given in GDUSHD definition andthen divided by 10.

GENE SILENCING: The interruption or suppression of the expression of agene at the level of transcription or translation.

GENOTYPE: Refers to the genetic mark-up or profile of a cell ororganism.

GIBERS=GIBBERELLA EAR ROT (PINK MOLD) (Gibberella zeae): A 1 to 9 visualrating indicating the resistance to Gibberella Ear Rot. A higher scoreindicates a higher resistance. Data are collected only when sufficientselection pressure exists in the experiment measured.

GIBROT=GIBBERELLA STALK ROT SCORE: Score of stalk rot severity due toGibberella (Gibberella zeae). Expressed as a 1 to 9 score with 9 beinghighly resistant. Data are collected only when sufficient selectionpressure exists in the experiment measured.

GLFSPT=GRAY LEAF SPOT (Cercospora zeae-maydis): A 1 to 9 visual ratingindicating the resistance to Gray Leaf Spot. A higher score indicates ahigher resistance. Data are collected only when sufficient selectionpressure exists in the experiment measured.

GOSWLT=GOSS' WILT (Corynebacterium nebraskense): A 1 to 9 visual ratingindicating the resistance to Goss' Wilt. A higher score indicates ahigher resistance. Data are collected only when sufficient selectionpressure exists in the experiment measured.

GRAIN TEXTURE: A visual rating used to indicate the appearance of maturegrain observed in the middle third of the uppermost ear when welldeveloped. Grain or seed with a hard grain texture is indicated asflint; grain or seed with a soft grain texture is indicted as dent.Medium grain or seed texture may be indicated as flint-dent orintermediate. Other grain textures include flint-like, dent-like, sweet,pop, waxy and flour.

GRNAPP=GRAIN APPEARANCE: This is a 1 to 9 rating for the generalappearance of the shelled grain as it is harvested based on such factorsas the color of harvested grain, any mold on the grain, and any crackedgrain. Higher scores indicate better grain visual quality.

H and H1: Refers to the haploid generation.

HAPLOID PLANT PART: Refers to a plant part or cell that has a haploidgenotype.

HCBLT=HELMINTHOSPORIUM CARBONUM LEAF BLIGHT (Helminthosporium carbonum):A 1 to 9 visual rating indicating the resistance to Helminthosporiuminfection. A higher score indicates a higher resistance. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

HD SMT=HEAD SMUT (Sphacelotheca reiliana): This indicates the percentageof plants not infected. Data are collected only when sufficientselection pressure exists in the experiment measured.

HSKCVR=HUSK COVER: A 1 to 9 score based on performance relative to keychecks, with a score of 1 indicating very short husks, tip of ear andkernels showing; 5 is intermediate coverage of the ear under mostconditions, sometimes with thin husk; and a 9 has husks extending andclosed beyond the tip of the ear. Scoring can best be done nearphysiological maturity stage or any time during dry down untilharvested.

HTFRM=Near-infrared transmission spectroscopy, NIT: prediction offermentables.

HYBRID VARIETY: A substantially heterozygous hybrid line and minorgenetic modifications thereof that retain the overall genetics of thehybrid line including but not limited to a locus conversion, a mutation,or a somoclonal variant.

INBRED: A variety developed through inbreeding or doubled haploidy thatpreferably comprises homozygous alleles at about 95% or more of itsloci. An inbred can be reproduced by selfing or growing in isolation sothat the plants can only pollinate with the same inbred variety.

INC D/A=GROSS INCOME (DOLLARS PER ACRE): Relative income per acreassuming drying costs of two cents per point above 15.5 percent harvestmoisture and current market price per bushel.

INCOME/ACRE: Income advantage of hybrid to be patented over other hybridon per acre basis.

INC ADV=GROSS INCOME ADVANTAGE: Gross income advantage of variety #1over variety #2.

INTROGRESSION: The process of transferring genetic material from onegenotype to another.

KERUNT=KERNELS PER UNIT AREA (Acres or Hectares).

KERPOP=KERNEL POP SCORE: The visual 1-9 rating of the amount ofrupturing of the kernel pericarp at an early stage in grain fill. Ahigher score indicates fewer popped (ruptured) kernels.

KER_WT=KERNEL NUMBER PER UNIT WEIGHT (Pounds or Kilograms): The numberof kernels in a specific measured weight; determined after removal ofextremely small and large kernels.

KSZDCD=KERNEL SIZE DISCARD: The percent of discard seed; calculated asthe sum of discarded tip kernels and extra-large kernels.

LINKAGE: Refers to a phenomenon wherein alleles on the same chromosometend to segregate together more often than expected by chance if theirtransmission was independent.

LINKAGE DISEQUILIBRIUM: Refers to a phenomenon wherein alleles tend toremain together in linkage groups when segregating from parents tooffspring, with a greater frequency than expected from their individualfrequencies.

LOCUS: A specific location on a chromosome.

LOCUS CONVERSION (Also called TRAIT CONVERSION): A locus conversionrefers to plants within a variety that have been modified in a mannerthat retains the overall genetics of the variety and further comprisesone or more loci with a specific desired trait, such as male sterility,insect resistance, disease resistance or herbicide tolerance orresistance. Examples of single locus conversions include mutant genes,transgenes and native traits finely mapped to a single locus. One ormore locus conversion traits may be introduced into a single cornvariety.

L/POP=YIELD AT LOW DENSITY: Yield ability at relatively low plantdensities on a 1 to 9 relative system with a higher number indicatingthe hybrid responds well to low plant densities for yield relative toother hybrids. A 1, 5, and 9 would represent very poor, average, andvery good yield response, respectively, to low plant density.

LRTLDG=LATE ROOT LODGING: The percentage of plants that do not rootlodge after anthesis through harvest; plants that lean from the verticalaxis at an approximately 30 degree angle or greater would be counted asroot lodged. Data are collected only when sufficient selection pressureexists in the experiment measured.

LRTLPN=LATE ROOT LODGING: An estimate of the percentage of plants thatdo not root lodge after anthesis through harvest; plants that lean fromthe vertical axis at an approximately 30 degree angle or greater wouldbe considered as root lodged. Data are collected only when sufficientselection pressure exists in the experiment measured.

LRTLSC=LATE ROOT LODGING SCORE: Score for severity of plants that leanfrom a vertical axis at an approximate 30 degree angle or greater whichtypically results from strong winds after flowering. Recorded prior toharvest when a root-lodging event has occurred. This lodging results inplants that are leaned or “lodged” over at the base of the plant and donot straighten or “goose-neck” back to a vertical position. Expressed asa 1 to 9 score with 9 being no lodging. Data are collected only whensufficient selection pressure exists in the experiment measured.

MALE STERILITY: A male sterile plant is one which produces no viablepollen no (pollen that is able to fertilize the egg to produce a viableseed). Male sterility prevents self pollination. These male sterileplants are therefore useful in hybrid plant production.

MDMCPX=MAIZE DWARF MOSAIC COMPLEX (MDMV=Maize Dwarf Mosaic Virus andMCDV=Maize Chlorotic Dwarf Virus): A 1 to 9 visual rating indicating theresistance to Maize Dwarf Mosaic Complex. A higher score indicates ahigher resistance. Data are collected only when sufficient selectionpressure exists in the experiment measured.

MILKLN=percent milk in mature grain.

MST=HARVEST MOISTURE: The moisture is the actual percentage moisture ofthe grain at harvest.

MSTADV=MOISTURE ADVANTAGE: The moisture advantage of variety #1 overvariety #2 as calculated by: MOISTURE of variety #2−MOISTURE of variety#1=MOISTURE ADVANTAGE of variety #1.

NEI DISTANCE: A quantitative measure of percent similarity between twovarieties. Nei's distance between varieties A and B can be defined as1−(2*number alleles in common/(number alleles in A+number alleles in B).For example, if varieties A and B are the same for 95 out of 100alleles, the Nei distance would be 0.05. If varieties A and B are thesame for 98 out of 100 alleles, the Nei distance would be 0.02. Freesoftware for calculating Nei distance is available on the internet atmultiple locations. See Nei, Proc Natl Acad Sci, 76:5269-5273 (1979)which is incorporated by reference for this purpose.

NLFBLT=NORTHERN LEAF BLIGHT (Helminthosporium turcicum or Exserohilumturcicum): A 1 to 9 visual rating indicating the resistance to NorthernLeaf Blight. A higher score indicates a higher resistance. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

NUCLEIC ACID: An acidic, chainlike biological macromolecule consistingof multiple repeat units of phosphoric acid, sugar, and purine andpyrimidine bases.

OILT=GRAIN OIL: Absolute value of oil content of the kernel as predictedby Near-Infrared Transmittance and expressed as a percent of dry matter.

PERCENT IDENTITY: Percent identity as used herein refers to thecomparison of the alleles present in two varieties. For example, whencomparing two inbred plants to each other, each inbred plant will havethe same allele (and therefore be homozygous) at almost all of theirloci. Percent identity is determined by comparing a statisticallysignificant number of the homozygous alleles of two varieties. Forexample, a percent identity of 90% between PH2FRV and other varietymeans that the two varieties have the same homozygous alleles at 90% oftheir loci.

PLANT: As used herein, the term “plant” includes reference to animmature or mature whole plant, including a plant that has beendetasseled or from which seed or grain has been removed. Seed or embryothat will produce the plant is also considered to be the plant.

PLANT PART: As used herein, the term “plant part” includes leaves,stems, roots, seed, grain, embryo, pollen, ovules, flowers, ears, cobs,husks, stalks, root tips, anthers, pericarp, silk, tissue, cells and thelike. In some embodiments, the plant part contains at least one cell ofinbred maize variety PH2FRV.

PLATFORM indicates the variety with the base genetics and the varietywith the base genetics comprising locus conversion(s). There can be aplatform for the inbred maize variety and the hybrid maize variety.

PLTHT=EDPLTHWT=PLANT HEIGHT: This is a measure of the height of theplant from the ground to the tip of the tassel in inches (PLTHT) orcentimeters (EDPLTHWT).

POLPRD=POLLEN PRODUCTION SCORE: The estimated total amount of pollenproduced by tassels based on the number of tassel branches and thedensity of the spikelets.

POLSC=POLLEN SCORE: A 0 to 9 visual rating indicating the amount ofpollen shed. The higher the score the more pollen shed.

POLWT=POLLEN WEIGHT: This is calculated by dry weight of tasselscollected as shedding commences minus dry weight from similar tasselsharvested after shedding is complete.

POP K/A=PLANT POPULATIONS: Measured as 1000's per acre.

POP ADV=PLANT POPULATION ADVANTAGE: The plant population advantage ofvariety #1 over variety #2 as calculated by PLANT POPULATION of variety#2−PLANT POPULATION of variety #1=PLANT POPULATION ADVANTAGE of variety#1.

RM=RELATIVE MATURITY: This is a predicted relative maturity, based onthe harvest moisture of the grain. The relative maturity rating is basedon a known set of checks and utilizes standard linear regressionanalyses and is also referred to as the Comparative Relative MaturityRating System that is similar to the Minnesota Relative Maturity RatingSystem.

PRMSHD: A relative measure of the growing degree units (GDU) required toreach 50% pollen shed. Relative values are predicted values from thelinear regression of observed GDU's on relative maturity of commercialchecks.

PROT=GRAIN PROTEIN: Absolute value of protein content of the kernel aspredicted by Near-Infrared Transmittance and expressed as a percent ofdry matter.

RESISTANCE: Synonymous with tolerance. The ability of a plant towithstand exposure to an insect, disease, herbicide or other condition.A resistant plant variety will have a level of resistance higher than acomparable wild-type variety.

RTLDG=ROOT LODGING: Root lodging is the percentage of plants that do notroot lodge; plants that lean from the vertical axis at an approximately30 degree angle or greater would be counted as root lodged. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

RTLADV=ROOT LODGING ADVANTAGE: The root lodging advantage of variety #1over variety #2. Data are collected only when sufficient selectionpressure exists in the experiment measured.

SCTGRN=SCATTER GRAIN: A 1 to 9 visual rating indicating the amount ofscatter grain (lack of pollination or kernel abortion) on the ear. Thehigher the score the less scatter grain.

SDGVGR=SEEDLING VIGOR: This is the visual rating (1 to 9) of the amountof vegetative growth after emergence at the seedling stage(approximately five leaves). A higher score indicates better vigor.

SEED: Fertilized and ripened ovule, consisting of the plant embryo,varying amounts of stored food material, and a protective outer seedcoat. Synonymous with grain.

SEFIELD: Percent stress emergence in field.

SELAB: Average % stress emergence in lab tests.

SEL IND=SELECTION INDEX: The selection index gives a single measure ofthe hybrid's worth based on information for multiple traits. A maizebreeder may utilize his or her own set of traits for the selectionindex. One of the traits that is almost always included is yield. Theselection index data presented in the tables represent the mean valueaveraged across testing stations.

SELF POLLINATION: A plant is self-pollinated if pollen from one floweris transferred to the same or another flower of the same plant.

SIB POLLINATION: A plant is sib-pollinated when individuals within thesame family or variety are used for pollination.

SITE SPECIFIC INTEGRATION: Genes that create a site for site specificDNA integration. This includes the introduction of FRT sites that may beused in the FLP/FRT system and/or Lox sites that may be used in theCre/Loxp system. For example, see Lyznik, et al., Site-SpecificRecombination for Genetic Engineering in Plants, Plant Cell Rep (2003)21:925-932 and WO 99/25821.

SLFBLT=SOUTHERN LEAF BLIGHT (Helminthosporium maydis or Bipolarismaydis): A 1 to 9 visual rating indicating the resistance to SouthernLeaf Blight. A higher score indicates a higher resistance. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

SNP=SINGLE-NUCLEOTIDE POLYMORPHISM: is a DNA sequence variationoccurring when a single nucleotide in the genome differs betweenindividual plant or plant varieties. The differences can be equated withdifferent alleles, and indicate polymorphisms. A number of SNP markerscan be used to determine a molecular profile of an individual plant orplant variety and can be used to compare similarities and differencesamong plants and plant varieties.

SOURST=SOUTHERN RUST (Puccinia polysora): A 1 to 9 visual ratingindicating the resistance to Southern Rust. A higher score indicates ahigher resistance. Data are collected only when sufficient selectionpressure exists in the experiment measured.

SPKDSC=EDTASAFDs=TASSEL SPIKELET DENSITY SCORE: The visual rating of howdense spikelets are on the middle to middle third of tassel branches. Ahigher score on a 1-9 scale indicates higher spikelet density (SPKDSC).On a 3 to 7 scale, 3 is moderately lax, 5 is medium, and 7 is moderatelydense (EDTASAFDs).

STAGRN=STAY GREEN: Stay green is the measure of plant health near thetime of black layer formation (physiological maturity). A high scoreindicates better late-season plant health.

STDADV=STALK STANDING ADVANTAGE: The advantage of variety #1 overvariety #2 for the trait STKCNT.

STKCNT=NUMBER OF PLANTS: This is the final stand or number of plants perplot.

STKCTE: This is the early stand count of plants per plot.

STKLDG=STALK LODGING REGULAR: This is the percentage of plants that didnot stalk lodge (stalk breakage) at regular harvest (when grain moistureis between about 20% and 30%) as measured by either natural lodging orpushing the stalks and determining the percentage of plants that breakbelow the ear. Data are collected only when sufficient selectionpressure exists in the experiment measured.

STKLDS=STALK LODGING SCORE: A plant is considered as stalk lodged if thestalk is broken or crimped between the ear and the ground. This can becaused by any or a combination of the following: strong winds late inthe season, disease pressure within the stalks, ECB damage orgenetically weak stalks. This trait should be taken just prior to or atharvest. Expressed on a 1 to 9 scale with 9 being no lodging. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

STLLPN=LATE STALK LODGING: This is the percent of plants that did notstalk lodge (stalk breakage or crimping) at or around late seasonharvest (when grain moisture is below 20%) as measured by either naturallodging or pushing the stalks and determining the percentage of plantsthat break or crimp below the ear. Data are collected only whensufficient selection pressure exists in the experiment measured.

STLPCN=STALK LODGING REGULAR: This is an estimate of the percentage ofplants that did not stalk lodge (stalk breakage) at regular harvest(when grain moisture is between about 20% and 30%) as measured by eithernatural lodging or pushing the stalks and determining the percentage ofplants that break below the ear. Data are collected only when sufficientselection pressure exists in the experiment measured.

STLTIP=STERILE TIPS SCORE: The visual 1 to 9 rating of the relative lackof glumes on the tassel central spike and branches. A higher scoreindicates less incidence of sterile tips or lack of glumes on thetassel.

STRT=GRAIN STARCH: Absolute value of starch content of the kernel aspredicted by Near-Infrared Transmittance and expressed as a percent ofdry matter.

STWWLT=Stewart's Wilt (Erwinia stewartii): A 1 to 9 visual ratingindicating the resistance to Stewart's Wilt. A higher score indicates ahigher resistance. Data are collected only when sufficient selectionpressure exists in the experiment measured.

SSRs: Genetic markers based on polymorphisms in repeated nucleotidesequences, such as microsatellites. A marker system based on SSRs can behighly informative in linkage analysis relative to other marker systemsin that multiple alleles may be present.

TASBLS=TASSEL BLAST: A 1 to 9 visual rating was used to measure thedegree of blasting (necrosis due to heat stress) of the tassel at thetime of flowering. A 1 would indicate a very high level of blasting attime of flowering, while a 9 would have no tassel blasting. Data arecollected only when sufficient selection pressure exists in theexperiment measured.

TASBRN=TASSEL BRANCH NUMBER: The number of tassel branches, with anthersoriginating from the central spike.

TASSZ=TASSEL SIZE: A 1 to 9 visual rating was used to indicate therelative size of the tassel. A higher rating means a larger tassel.

TAS WT=TASSEL WEIGHT: This is the average weight of a tassel (grams)just prior to pollen shed.

TILLER=TILLERS: A count of the number of tillers per plot that couldpossibly shed pollen was taken. Data are given as a percentage oftillers: number of tillers per plot divided by number of plants perplot. A tiller is defined as a secondary shoot that has developed as atassel capable of shedding pollen.

TSTWT=TEST WEIGHT (ADJUSTED): The measure of the weight of the grain inpounds for a given volume (bushel), adjusted for MST less than or equalto 22%.

TSTWTN=TEST WEIGHT (UNADJUSTED): The measure of the weight of the grainin pounds for a given volume (bushel).

TSWADV=TEST WEIGHT ADVANTAGE: The test weight advantage of variety #1over variety #2.

VARIETY: A maize line and minor genetic modifications thereof thatretain the overall genetics of the line including but not limited to alocus conversion, a mutation, or a somoclonal variant.

WIN M %=PERCENT MOISTURE WINS.

WIN Y %=PERCENT YIELD WINS.

YIELD BU/A=YIELD (BUSHELS/ACRE): Yield of the grain at harvest by weightor volume (bushels) per unit area (acre) adjusted to 15% moisture.

YLDADV=YIELD ADVANTAGE: The yield advantage of variety #1 over variety#2 as calculated by: YIELD of variety #1−YIELD variety #2=YIELDADVANTAGE of variety #1.

YIELDMST=YIELD/MOISTURE RATIO.

YLDSC=YIELD SCORE: A 1 to 9 visual rating was used to give a relativerating for yield based on plot ear piles. The higher the rating thegreater visual yield appearance.

YIELDS=Silage Dry Matter Yield (tons/acre@ 100% DM)

MLKYLD=Estimated pounds of milk produced per ton of dry matter fed andis based on utilizing nutrient content and fiber digestibility

ADJMLK=Estimated pounds of milk produced per acre of silage dry matterbased on an equation and is MLKYLD divided by YIELDS.

SLGPRM=Silage Predicted Relative Maturity

SY30DM=Silage Yield (Tonnage) Adjusted to 30% Dry Matter

PCTMST=Silage Harvest Moisture %

NDFDR=Silage Fiber Digestibility Based on rumen fluid NIRS calibration

NDFDC=Silage Fiber Digestibility Based on rumen fluid NIRS calibration

All tables discussed in the Detailed Description section can found atthe end of the section.

Breeding History of PH2FRV

Inbred Maize variety PH2FRV was developed by the following method. Across was made between inbred line PH12W6 and inbred line PH17M5. InbredPH2FRV was developed by producing a doubled haploid from the F1 plants,selfing and using pedigree selection amongst the D1 lines, and selfingand bulking from the subsequent generations.

Maize variety PH2FRV, being substantially homozygous, can be reproducedby planting seeds of the variety, growing the resulting maize plantsunder self-pollinating or sib-pollinating conditions with adequateisolation, and harvesting the resulting seed using techniques familiarto the agricultural arts.

Phenotypic Characteristics of PH2FRV

Inbred maize variety PH2FRV may be used as a male or female in theproduction of the first generation F1 hybrid. The variety has shownuniformity and stability within the limits of environmental influencefor all the traits as described in the Variety Description Information(Table 1, found at the end of the section). The variety has beenself-pollinated and ear-rowed a sufficient number of generations withcareful attention paid to uniformity of plant type to ensure sufficienthomozygosity and phenotypic stability for use in commercial hybrid seedproduction. The variety has been increased both by hand and in isolatedfields with continued observation for uniformity. No variant traits havebeen observed or are expected in PH2FRV.

Genotypic Characteristics of PH2FRV

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile.

As a result of inbreeding, PH2FRV is substantially homozygous. Thishomozygosity can be characterized at the loci shown in a marker profile.An F1 hybrid made with PH2FRV would substantially comprise the markerprofile of PH2FRV. This is because an F1 hybrid is the sum of its inbredparents, e.g., if one inbred parent is homozygous for allele x at aparticular locus, and the other inbred parent is homozygous for allele yat that locus, the F1 hybrid will be xy (heterozygous) at that locus. Agenetic marker profile can therefore be used to identify hybridscomprising PH2FRV as a parent, since such hybrids will comprise two setsof alleles, one set of which will be from PH2FRV. The determination ofthe male set of alleles and the female set of alleles may be made byprofiling the hybrid and the pericarp of the hybrid seed, which iscomposed of maternal parent cells. One way to obtain the paternal parentprofile is to subtract the pericarp profile from the hybrid profile.

Subsequent generations of progeny produced by selection and breeding areexpected to be of genotype xx (homozygous), yy (homozygous), or xy(heterozygous) for these locus positions. When the F1 plant is used toproduce an inbred, the resulting inbred should be either x or y for thatallele.

Therefore, in accordance with the above, an embodiment is a PH2FRVprogeny maize plant or plant part that is a first generation (F1) hybridmaize plant comprising two sets of alleles, wherein one set of thealleles is the same as PH2FRV at substantially all loci. A maize cellwherein one set of the alleles is the same as PH2FRV at substantiallyall loci is also an embodiment of the invention. This maize cell may bea part of a hybrid seed, plant or plant part produced by crossing PH2FRVwith another maize plant.

Genetic marker profiles can be obtained by techniques such asRestriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asMicrosatellites, and Single Nucleotide Polymorphisms (SNPs). Forexample, see Berry et al. (2002), “Assessing Probability of AncestryUsing Simple Sequence Repeat Profiles: Applications to Maize Hybrids andInbreds”, Genetics, 2002, 161:813-824, and Berry et al. (2003),“Assessing Probability of Ancestry Using Simple Sequence RepeatProfiles: Applications to Maize Inbred Lines and Soybean Varieties”,Genetics, 2003, 165: 331-342.

Particular markers used for these purposes may include any type ofmarker and marker profile which provides a means of distinguishingvarieties. A genetic marker profile can be used, for example, toidentify plants of the same variety or related varieties or to determineor validate a pedigree. In addition to being used for identification ofmaize variety PH2FRV, a hybrid produced through the use of PH2FRV, andthe identification or verification of pedigree for progeny plantsproduced through the use of PH2FRV, a genetic marker profile is alsouseful in developing a locus conversion of PH2FRV.

Methods of isolating nucleic acids from maize plants and methods forperforming genetic marker profiles using SNP and SSR polymorphisms arewell known in the art. SNPs are genetic markers based on a polymorphismin a single nucleotide. A marker system based on SNPs can be highlyinformative in linkage analysis relative to other marker systems in thatmultiple alleles may be present. A method comprising isolating nucleicacids, such as DNA, from a plant, a plant part, plant cell or a seed ofthe maize plants disclosed herein is provided. The method can includemechanical, electrical and/or chemical disruption of the plant, plantpart, plant cell or seed, contacting the disrupted plant, plant part,plant cell or seed with a buffer or solvent, to produce a solution orsuspension comprising nucleic acids, optionally contacting the nucleicacids with a precipiting agent to precipitate the nucleic acids,optionally extracting the nucleic acids, and optionally separating thenucleic acids such as by centrifugation or by binding to beads or acolumn, with subsequent elution, or a combination thereof. If DNA isbeing isolated, an RNase can be included in one or more of the methodsteps. The nucleic acids isolated can comprise all or substantially allof the genomic DNA sequence, all or substantially all of the chromosomalDNA sequence or all or substantially all of the coding sequences (cDNA)of the plant, plant part, or plant cell from which they were isolated.The nucleic acids isolated can comprise all, substantially all, oressentially all of the genetic complement of the plant. The amount andtype of nucleic acids isolated may be sufficient to permit whole genomesequencing of the plant from which they were isolated or chromosomalmarker analysis of the plant from which they were isolated.

The methods can be used to produce nucleic acids from the plant, plantpart, seed or cell, which nucleic acids can be, for example, analyzed toproduce data. The data can be recorded. The nucleic acids from thedisrupted cell, the disrupted plant, plant part, plant cell or seed orthe nucleic acids following isolation or separation can be contactedwith primers and nucleotide bases, and/or a polymerase to facilitate PCRsequencing or marker analysis of the nucleic acids. In some examples,the nucleic acids produced can be sequenced or contacted with markers toproduce a genetic profile, a molecular profile, a marker profile, ahaplotype, or any combination thereof. In some examples, the geneticprofile or nucleotide sequence is recorded on a computer readablemedium. In other examples, the methods may further comprise using thenucleic acids produced from plants, plant parts, plant cells or seeds ina plant breeding program, for example in making crosses, selectionand/or advancement decisions in a breeding program. Crossing includesany type of plant breeding crossing method, including but not limited tocrosses to produce hybrids, outcrossing, selfing, backcrossing, locusconversion, introgression and the like.

Favorable genotypes and or marker profiles, optionally associated with atrait of interest, may be identified by one or more methodologies. Insome examples one or more markers are used, including but not limited toAFLPs, RFLPs, ASH, SSRs, SNPs, indels, padlock probes, molecularinversion probes, microarrays, sequencing, and the like. In somemethods, a target nucleic acid is amplified prior to hybridization witha probe. In other cases, the target nucleic acid is not amplified priorto hybridization, such as methods using molecular inversion probes (see,for example Hardenbol et al. (2003) Nat Biotech 21:673-678). In someexamples, the genotype related to a specific trait is monitored, whilein other examples, a genome-wide evaluation including but not limited toone or more of marker panels, library screens, association studies,microarrays, gene chips, expression studies, or sequencing such aswhole-genome resequencing and genotyping-by-sequencing (GBS) may beused. In some examples, no target-specific probe is needed, for exampleby using sequencing technologies, including but not limited tonext-generation sequencing methods (see, for example, Metzker (2010) NatRev Genet 11:31-46; and, Egan et al. (2012) Am J Bot 99:175-185) such assequencing by synthesis (e.g., Roche 454 pyrosequencing, Illumina GenomeAnalyzer, and Ion Torrent PGM or Proton systems), sequencing by ligation(e.g., SOLiD from Applied Biosystems, and Polnator system from AzcoBiotech), and single molecule sequencing (SMS or third-generationsequencing) which eliminate template amplification (e.g., Helicossystem, and PacBio RS system from Pacific BioSciences). Furthertechnologies include optical sequencing systems (e.g., Starlight fromLife Technologies), and nanopore sequencing (e.g., GridION from OxfordNanopore Technologies). Each of these may be coupled with one or moreenrichment strategies for organellar or nuclear genomes in order toreduce the complexity of the genome under investigation via PCR,hybridization, restriction enzyme (see, e.g., Elshire et al. (2011) PLoSONE 6:e19379), and expression methods. In some examples, no referencegenome sequence is needed in order to complete the analysis. PH2FRV andits plant parts can be identified through a molecular marker profile.Such plant parts may be either diploid or haploid. Also encompassedwithin the scope of the invention are plants and plant partssubstantially benefiting from the use of variety PH2FRV in theirdevelopment, such as variety PH2FRV comprising a locus conversion.

Comparing PH2FRV to Other Inbreds

A breeder uses various methods to help determine which plants should beselected from segregating populations and ultimately which inbredvarieties will be used to develop hybrids for commercialization. Inaddition to knowledge of the germplasm and plant genetics, a part of theselection process is dependent on experimental design coupled with theuse of statistical analysis. Experimental design and statisticalanalysis are used to help determine which plants, which family ofplants, and finally which inbred varieties and hybrid combinations aresignificantly better or different for one or more traits of interest.Experimental design methods are used to assess error so that differencesbetween two inbred varieties or two hybrid varieties can be moreaccurately evaluated. Statistical analysis includes the calculation ofmean values, determination of the statistical significance of thesources of variation, and the calculation of the appropriate variancecomponents. Either a five or a one percent significance level iscustomarily used to determine whether a difference that occurs for agiven trait is real or due to the environment or experimental error. Oneof ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is asignificant difference between the two traits expressed by thosevarieties. For example, see Fehr, Walt, Principles of CultivarDevelopment, p. 261-286 (1987). Mean trait values may be used todetermine whether trait differences are significant. Trait values shouldpreferably be measured on plants grown under the same environmentalconditions, and environmental conditions should be appropriate for thetraits or traits being evaluated. Sufficient selection pressure shouldbe present for optimum measurement of traits of interest such asherbicide tolerance, insect or disease resistance. A locus conversion ofPH2FRV for herbicide tolerance should be compared with an isogeniccounterpart in the absence of the converted trait. In addition, a locusconversion for insect or disease resistance should be compared to theisogenic counterpart, in the absence of disease pressure or insectpressure.

Development of Maize Hybrids Using PH2FRV

A single cross maize hybrid results from the cross of two inbredvarieties, each of which has a genotype that complements the genotype ofthe other. A hybrid progeny of the first generation is designated F1. Inthe development of commercial hybrids in a maize plant breeding program,only the F1 hybrid plants are sought. F1 hybrids are more vigorous thantheir inbred parents. This hybrid vigor, or heterosis, can be manifestedin many polygenic traits, including increased vegetative growth andincreased yield.

PH2FRV may be used to produce hybrid maize. One such embodiment is themethod of crossing maize variety PH2FRV with another maize plant, suchas a different maize variety, to form a first generation F1 hybrid seed.The first generation F1 hybrid seed, plant and plant part produced bythis method is an embodiment of the invention. The first generation F1seed, plant and plant part will comprise an essentially complete set ofthe alleles of variety PH2FRV. One of ordinary skill in the art canutilize molecular methods to identify a particular F1 hybrid plantproduced using variety PH2FRV. Further, one of ordinary skill in the artmay also produce F1 hybrids with transgenic, male sterile and/or locusconversions of variety PH2FRV.

The development of a maize hybrid in a maize plant breeding programinvolves three steps: (1) the selection of plants from various germplasmpools for initial breeding crosses; (2) the selfing of the selectedplants from the breeding crosses for several generations to produce aseries of varieties, such as PH2FRV, which, although different from eachother, breed true and are highly uniform; and (3) crossing the selectedvarieties with different varieties to produce the hybrids. During theinbreeding process in maize, the vigor of the varieties decreases, andso one would not be likely to use PH2FRV directly to produce grain.However, vigor is restored when PH2FRV is crossed to a different inbredvariety to produce a commercial F1 hybrid. A consequence of thehomozygosity and homogeneity of the inbred variety is that the hybridbetween a defined pair of inbreds may be reproduced indefinitely as longas the homogeneity of the inbred parents is maintained.

PH2FRV may be used to produce a single cross hybrid, a double crosshybrid, or a three-way hybrid. A single cross hybrid is produced whentwo inbred varieties are crossed to produce the F1 progeny. A doublecross hybrid is produced from four inbred varieties crossed in pairs(A×B and C×D) and then the two F1 hybrids are crossed again (A×B)×(C×D).A three-way cross hybrid is produced from three inbred varieties wheretwo of the inbred varieties are crossed (A×B) and then the resulting F1hybrid is crossed with the third inbred (A×B)×C. In each case, pericarptissue from the female parent will be a part of and protect the hybridseed.

Molecular data from PH2FRV may be used in a plant breeding process.Nucleic acids may be isolated from a seed of PH2FRV or from a plant,plant part, or cell produced by growing a seed of PH2FRV, or from a seedof PH2FRV with a locus conversion, or from a plant, plant part, or cellof PH2FRV with a locus conversion. One or more polymorphisms may beisolated from the nucleic acids. A plant having one or more of theidentified polymorphisms may be selected and used in a plant breedingmethod to produce another plant.

Combining Ability of PH2FRV

Combining ability of a variety, as well as the performance of thevariety per se, is a factor in the selection of improved maize inbreds.Combining ability refers to a variety's contribution as a parent whencrossed with other varieties to form hybrids. The hybrids formed for thepurpose of selecting superior varieties may be referred to as testcrosses, and include comparisons to other hybrid varieties grown in thesame environment (same cross, location and time of planting). One way ofmeasuring combining ability is by using values based in part on theoverall mean of a number of test crosses weighted by number ofexperiment and location combinations in which the hybrid combinationsoccurs. The mean may be adjusted to remove environmental effects andknown genetic relationships among the varieties.

General combining ability provides an overall score for the inbred overa large number of test crosses. Specific combining ability providesinformation on hybrid combinations formed by PH2FRV and a specificinbred parent. A variety such as PH2FRV which exhibits good generalcombining ability may be used in a large number of hybrid combinations.

A general combining ability report for PH2FRV is provided in Table 2. InTable 2, found at the end of this section, BLUP, Best Linear UnbiasedPrediction, values are reported for the breeding value of the maizeinbred PH2FRV platform. The BLUP values are reported for numerous traitsof hybrids that have inbred PH2FRV or a locus conversion of PH2FRV as aparent. The inbred PH2FRV and various locus conversions of PH2FRV aretogether considered a platform. The values reported indicate a BLUPvalue averaged for all members of the platform weighted by the inverseof the Standard Errors.

Hybrid Comparisons

These hybrid comparisons represent specific hybrid crosses with PH2FRVand a comparison of these specific hybrids with other hybrids withfavorable characteristics. These comparisons illustrate the goodspecific combining ability of PH2FRV.

The results in Table 3 compare a specific hybrid for which PH2FRV is aparent with other hybrids. The data in Table 3 shows that numerousspecies of the genus of F1 hybrids created with PH2FRV have been reducedto practice. These comparisons illustrate the good specific combiningability of PH2FRV. In Table 3, found at the end of this section, BLUPvalues are reported for different hybrids wherein one parent is themaize variety PH2FRV or PH2FRV comprising locus conversions. The BLUPvalues and Standard Errors, SE, are reported for numerous traits. Thedata presented for these hybrids is based on replicated field trials.

Introduction of a New Trait or Locus into PH2FRV

Inbred PH2FRV represents a new base genetic variety into which a newlocus or trait may be introduced or introgressed. Transformation andbackcrossing represent two methods that can be used to accomplish suchan introgression. The term locus conversion is used to designate theproduct of such an introgression.

A backcross or locus conversion of PH2FRV occurs when DNA sequences areintroduced through backcrossing (Hallauer et al. in Corn and CornImprovement, Sprague and Dudley, Third Ed. 1998), with PH2FRV utilizedas the recurrent parent. Both naturally occurring, modified andtransgenic DNA sequences may be introduced through backcrossingtechniques. A backcross or locus conversion may produce a plant with atrait or locus conversion in at least one or more backcrosses, includingat least 2 backcrosses, at least 3 backcrosses, at least 4 backcrosses,at least 5 backcrosses and the like. Molecular marker assisted breedingor selection may be utilized to reduce the number of backcrossesnecessary to achieve the backcross conversion. For example, see Openshawet al., “Marker-assisted Selection in Backcross Breeding,” in:Proceedings Symposium of the Analysis of Molecular Data, August 1994,Crop Science Society of America, Corvallis, Oreg., which demonstratedthat a backcross locus conversion can be made in as few as twobackcrosses.

The complexity of the backcross conversion method depends on the type oftrait being transferred (a single gene or closely linked genes comparedto unlinked genes), the level of expression of the trait, the type ofinheritance (cytoplasmic or nuclear), dominant or recessive traitexpression, and the types of parents included in the cross. It isunderstood by those of ordinary skill in the art that for single locusor gene traits that are relatively easy to classify, the backcrossmethod is effective and relatively easy to manage. (See Hallauer et al.in Corn and Corn Improvement, Sprague and Dudley, Third Ed. 1998).Desired traits that may be transferred through backcross conversioninclude, but are not limited to, waxy starch, sterility (nuclear andcytoplasmic), fertility restoration, grain color (white), nutritionalenhancements, drought tolerance, nitrogen utilization, altered fattyacid profile, increased digestibility, low phytate, industrialenhancements, disease resistance (bacterial, fungal, or viral), insectresistance, and herbicide tolerance or resistance. A locus conversion,also called a trait conversion, can be a native trait or a transgenictrait. In addition, a recombination site itself, such as an FRT site,Lox site, or other site specific integration site may be inserted bybackcrossing and utilized for direct insertion of one or more genes ofinterest into a specific plant variety. A single locus may containseveral transgenes, such as a transgene for disease resistance that, inthe same expression vector, also contains a transgene for herbicidetolerance or resistance. The gene for herbicide tolerance or resistancemay be used as a selectable marker and/or as a phenotypic trait. Asingle locus conversion of a site specific integration system allows forthe integration of multiple genes at a known recombination site in thegenome. At least one, at least two or at least three and less than ten,less than nine, less than eight, less than seven, less than six, lessthan five or less than four locus conversions may be introduced into theplant by backcrossing, introgression or transformation to express thedesired trait, while the plant, or a plant grown from the seed, plantpart or plant cell, otherwise retains the phenotypic characteristics ofthe deposited seed when grown under the same environmental conditions.

The backcross or locus conversion may result from either the transfer ofa dominant allele or a recessive allele. Selection of progeny containingthe trait of interest can be accomplished by direct selection for atrait associated with a dominant allele. Transgenes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele, such as the waxy starchcharacteristic, requires growing and selfing the first backcrossgeneration to determine which plants carry the recessive alleles.Recessive traits may require additional progeny testing in successivebackcross generations to determine the presence of the locus ofinterest. The last backcross generation is usually selfed to give purebreeding progeny for the gene(s) being transferred, although a backcrossconversion with a stably introgressed trait may also be maintained byfurther backcrossing to the recurrent parent with selection for theconverted trait.

Along with selection for the trait of interest, progeny are selected forthe phenotype and/or genotype of the recurrent parent. Whileoccasionally additional polynucleotide sequences or genes may betransferred along with the backcross conversion, the backcrossconversion variety “fits into the same hybrid combination as therecurrent parent inbred variety and contributes the effect of theadditional locus added through the backcross.” Poehlman et al (1995)Breeding Field Crop, 4th Ed., Iowa State University Press, Ames, Iowa,pp. 132-155 and 321-344. When one or more traits are introgressed intothe variety a difference in quantitative agronomic traits, such as yieldor dry down, between the variety and an introgressed version of thevariety in some environments may occur. For example, the introgressedversion may provide a net yield increase in environments where the traitprovides a benefit, such as when a variety with an introgressed traitfor insect resistance is grown in an environment where insect pressureexists, or when a variety with herbicide tolerance is grown in anenvironment where herbicide is used.

One process for adding or modifying a trait or locus in maize varietyPH2FRV comprises crossing PH2FRV plants grown from PH2FRV seed withplants of another maize variety that comprise the desired trait orlocus, selecting F1 progeny plants that comprise the desired trait orlocus to produce selected F1 progeny plants, crossing the selectedprogeny plants with the PH2FRV plants to produce backcross progenyplants, selecting for backcross progeny plants that have the desiredtrait or locus and the phenotypic characteristics of maize varietyPH2FRV to produce selected backcross progeny plants; and backcrossing toPH2FRV one or more times in succession to produce backcross progenyplants that comprise said trait or locus. The modified PH2FRV may befurther characterized as having essentially the same phenotypiccharacteristics of maize variety PH2FRV listed in Table 1 and/or may becharacterized by percent identity to PH2FRV as determined by molecularmarkers, such as SSR markers or SNP markers.

In addition, the above process and other similar processes describedherein may be used to produce F1 hybrid maize seed by adding a step atthe end of the process that comprises crossing PH2FRV with the locusconversion with a different maize plant and harvesting the resultant F1hybrid maize seed.

Traits are also used by those of ordinary skill in the art tocharacterize progeny. Traits are commonly evaluated at a significancelevel, such as a 1%, 5% or 10% significance level, when measured inplants grown in the same environmental conditions.

Male Sterility and Hybrid Seed Production

Hybrid seed production requires elimination or inactivation of pollenproduced by the female inbred parent. Incomplete removal or inactivationof the pollen provides the potential for self-pollination. A reliablemethod of controlling male fertility in plants offers the opportunityfor improved seed production.

PH2FRV can be produced in a male-sterile form. There are several ways inwhich a maize plant can be manipulated so that it is male sterile. Theseinclude use of manual or mechanical emasculation (or detasseling), useof one or more genetic factors that confer male sterility, includingcytoplasmic genetic and/or nuclear genetic male sterility, use ofgametocides and the like. A male sterile designated PH2FRV may includeone or more genetic factors, which result in cytoplasmic genetic and/ornuclear genetic male sterility. All of such embodiments are within thescope of the present claims. The male sterility may be either partial orcomplete male sterility.

Hybrid maize seed is often produced by a male sterility systemincorporating manual or mechanical detasseling. Alternate strips of twomaize inbreds are planted in a field, and the pollen-bearing tassels areremoved from one of the inbreds (female). Provided that there issufficient isolation from sources of foreign maize pollen, the ears ofthe detasseled inbred will be fertilized only from the other inbred(male), and the resulting seed is therefore hybrid and will form hybridplants.

The laborious detasseling process can be avoided by using cytoplasmicmale-sterile (CMS) inbreds. Plants of a CMS inbred are male sterile as aresult of genetic factors in the cytoplasm, as opposed to the nucleus,and so nuclear linked genes are not transferred during backcrossing.Thus, this characteristic is inherited exclusively through the femaleparent in maize plants, since only the female provides cytoplasm to thefertilized seed. CMS plants are fertilized with pollen from anotherinbred that is not male-sterile. Pollen from the second inbred may ormay not contribute genes that make the hybrid plants male-fertile, andeither option may be preferred depending on the intended use of thehybrid. The same hybrid seed, a portion produced from detasseled fertilemaize and a portion produced using the CMS system, can be blended toinsure that adequate pollen loads are available for fertilization whenthe hybrid plants are grown. CMS systems have been successfully usedsince the 1950's, and the male sterility trait is routinely backcrossedinto inbred varieties. See Wych, Robert D. (1988) “Production of HybridSeed”, Corn and Corn Improvement, Ch. 9, pp. 565-607.

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar et al. and chromosomal translocations asdescribed by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Inaddition to these methods, Albertsen et al., U.S. Pat. No. 5,432,068,describes a system of nuclear male sterility which includes: identifyinga gene which is needed for male fertility; silencing this native genewhich is needed for male fertility; removing the native promoter fromthe essential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing, or turning “on”,the promoter, which in turn allows the gene that confers male fertilityto be transcribed.

These, and the other methods of conferring genetic male sterility in theart, each possess their own benefits and drawbacks. Some other methodsuse a variety of approaches such as delivering into the plant a geneencoding a cytotoxic substance associated with a male tissue specificpromoter or an antisense system in which a gene needed for fertility isidentified and an antisense to that gene is inserted in the plant (seeFabinjanski, et al. EPO 89/3010153.8 publication no. 329,308 and PCTapplication PCT/CA90/00037 published as WO 90/08828).

Another system for controlling male sterility makes use of gametocides.Gametocides are not a genetic system, but rather a topical applicationof chemicals. These chemicals affect cells that are needed for malefertility. The application of these chemicals affects fertility in theplants only for the growing season in which the gametocide is applied(see Carlson, Glenn R., and U.S. Pat. No. 4,936,904). Application of thegametocide, timing of the application and genotype specificity oftenlimit the usefulness of the approach and it is not appropriate in allsituations.

Incomplete control over male fertility may result in self-pollinatedseed being unintentionally harvested and packaged with hybrid seed. Thiswould typically be only female parent seed, because the male plant isgrown in rows that are typically destroyed prior to seed development.Once the seed from the hybrid bag is planted, it is possible to identifyand select these self-pollinated plants. These self-pollinated plantswill be one of the inbred varieties used to produce the hybrid. Thoughthe possibility of PH2FRV being included in a hybrid seed bag exists,the occurrence is very low because much care is taken by seed companiesto avoid such inclusions. It is worth noting that hybrid seed is sold togrowers for the production of grain or forage and not for breeding orseed production. These self-pollinated plants can be identified andselected by one skilled in the art due to their less vigorous appearancefor vegetative and/or reproductive characteristics, including shorterplant height, small ear size, ear and kernel shape, or othercharacteristics.

Identification of these self-pollinated varieties can also beaccomplished through molecular marker analyses. See, “The Identificationof Female Selfs in Hybrid Maize: A Comparison Using Electrophoresis andMorphology”, Smith, J. S. C. and Wych, R. D., Seed Science andTechnology 14, 1-8 (1995), the disclosure of which is expresslyincorporated herein by reference. Through these technologies, thehomozygosity of the self-pollinated variety can be verified by analyzingallelic composition at various loci along the genome. Those methodsallow for rapid identification of the plants disclosed herein. See also,“Identification of Atypical Plants in Hybrid Maize Seed by Postcontroland Electrophoresis” Sarca, V. et al., Probleme de Genetica Teoritica siAplicata Vol. 20 (1) p. 29-42.

Transformation

Transgenes and transformation methods facilitate engineering of thegenome of plants to contain and express heterologous genetic elements,such as foreign genetic elements, or additional copies of endogenouselements, or modified versions of native or endogenous genetic elementsin order to alter at least one trait of a plant in a specific manner.Any sequences, such as DNA, whether from a different species or from thesame species, which have been stably inserted into a genome usingtransformation are referred to herein collectively as “transgenes”and/or “transgenic events”. Transgenes can be moved from one genome toanother using breeding techniques which may include, for example,crossing, backcrossing or double haploid production. In someembodiments, a transformed variant of PH2FRV may comprise at least onetransgene but could contain at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10and/or no more than 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2.Transformed versions of the claimed maize variety PH2FRV as well ashybrid combinations containing and inheriting the transgene thereof areprovided. F1 hybrid seed are provided which are produced by crossing adifferent maize plant with maize variety PH2FRV comprising a transgenewhich has been introduced into maize variety PH2FRV by backcrossing orgenetic transformation and is inherited by the F1 hybrid seed.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glick,B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88 and Armstrong, “The First Decade of Maize Transformation: A Reviewand Future Perspective” (Maydica 44:101-109, 1999). In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. See, forexample, Gruber et al., “Vectors for Plant Transformation” in Methods inPlant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J.E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89-119.

In general, methods to transform, modify, edit or alter plant endogenousgenomic DNA include altering the plant native DNA sequence or apre-existing transgenic sequence including regulatory elements, codingand non-coding sequences. These methods can be used, for example, totarget nucleic acids to pre-engineered target recognition sequences inthe genome. Such pre-engineered target sequences may be introduced bygenome editing or modification. As an example, a genetically modifiedplant variety is generated using “custom” or engineered endonucleasessuch as meganucleases produced to modify plant genomes (see e.g., WO2009/114321; Gao et al. (2010) Plant Journal 1:176-187). Anothersite-directed engineering method is through the use of zinc fingerdomain recognition coupled with the restriction properties ofrestriction enzyme. See e.g., Urnov, et al., (2010) Nat Rev Genet.11(9):636-46; Shukla, et al., (2009) Nature 459 (7245):437-41. Atranscription activator-like (TAL) effector-DNA modifying enzyme (TALEor TALEN) is also used to engineer changes in plant genome. See e.g.,US20110145940, Cermak et al., (2011) Nucleic Acids Res. 39(12) and Bochet al., (2009), Science 326(5959): 1509-12. Site-specific modificationof plant genomes can also be performed using the bacterial type IICRISPR (clustered regularly interspaced short palindromic repeats)/Cas(CRISPR-associated) system. See e.g., Belhaj et al., (2013), PlantMethods 9: 39; The Cas9/guide RNA-based system allows targeted cleavageof genomic DNA guided by a customizable small noncoding RNA in plants(see e.g., WO 2015026883A1, incorporated herein by reference).

Plant transformation methods may involve the construction of anexpression vector. Such a vector comprises a DNA sequence that containsa gene under the control of or operatively linked to a regulatoryelement, for example a promoter. The vector may contain one or moregenes and one or more regulatory elements.

A transgenic event which has been stably engineered into the germ cellline of a particular maize plant using transformation techniques, couldbe moved into the germ cell line of another variety using traditionalbreeding techniques that are well known in the plant breeding arts. Forexample, a backcrossing approach is commonly used to move a transgenicevent from a transformed maize plant to another variety, and theresulting progeny would then comprise the transgenic event(s). Also, ifan inbred variety was used for the transformation then the transgenicplants could be crossed to a different inbred in order to produce atransgenic hybrid maize plant.

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include, but are not limited to genes;coding sequences; inducible, constitutive, and tissue specificpromoters; enhancing sequences; and signal and targeting sequences. Forexample, see the traits, genes and transformation methods listed in U.S.Pat. Nos. 6,118,055 and 6,284,953, which are herein incorporated byreference. In addition, transformability of a variety can be increasedby introgressing the trait of high transformability from another varietyknown to have high transformability, such as Hi-II. See U.S. PatentApplication Publication US2004/0016030 (2004).

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants that areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem. 114: 92-6(1981).

Transgenic events can be mapped by one of ordinary skill in the art andsuch techniques are well known to those of ordinary skill in the art.For exemplary methodologies in this regard, see for example, Glick andThompson, Methods in Plant Molecular Biology and Biotechnology 269-284(CRC Press, Boca Raton, 1993).

Likewise, by means of the present invention, plants can be geneticallyengineered to express various phenotypes of agronomic interest. Throughthe transformation of maize the expression of genes can be altered toenhance disease resistance, insect resistance, herbicide tolerance,agronomic traits, grain quality and other traits. Transformation canalso be used to insert DNA sequences which control or help controlmale-sterility. DNA sequences native to maize as well as non-native DNAsequences can be transformed into maize and used to alter levels ofnative or non-native proteins. Various promoters, targeting sequences,enhancing sequences, and other DNA sequences can be inserted into themaize genome for the purpose of altering the expression of proteins.Reduction of the activity of specific genes (also known as genesilencing, or gene suppression) is desirable for several aspects ofgenetic engineering in plants.

Many techniques for gene silencing are well known to one of skill in theart, including but not limited to knock-outs (such as by insertion of atransposable element such as mu (Vicki Chandler, The Maize Handbook Ch.118 (Springer-Verlag 1994) or other genetic elements such as a FRT, Loxor other site specific integration site, antisense technology (see,e.g., Sheehy et al. (1988) PNAS USA 85:8805-8809; and U.S. Pat. Nos.5,107,065; 5,453,566; and 5,759,829); co-suppression (e.g., Taylor(1997) Plant Cell 9:1245; Jorgensen (1990) Trends Biotech.8(12):340-344; Flavell (1994) PNAS USA 91:3490-3496; Finnegan et al.(1994) Bio/Technology 12: 883-888; and Neuhuber et al. (1994) Mol. Gen.Genet. 244:230-241); RNA interference (Napoli et al. (1990) Plant Cell2:279-289; U.S. Pat. No. 5,034,323; Sharp (1999) Genes Dev. 13:139-141;Zamore et al. (2000) Cell 101:25-33; and Montgomery et al. (1998) PNASUSA 95:15502-15507), virus-induced gene silencing (Burton, et al. (2000)Plant Cell 12:691-705; and Baulcombe (1999) Curr. Op. Plant Bio.2:109-113); target-RNA-specific ribozymes (Haseloff et al. (1988) Nature334: 585-591); hairpin structures (Smith et al. (2000) Nature407:319-320; WO 99/53050; and WO 98/53083); MicroRNA (Aukerman & Sakai(2003) Plant Cell 15:2730-2741); ribozymes (Steinecke et al. (1992) EMBOJ. 11:1525; and Perriman et al. (1993) Antisense Res. Dev. 3:253);oligonucleotide mediated targeted modification (e.g., WO 03/076574 andWO 99/25853); Zn-finger targeted molecules (e.g., WO 01/52620; WO03/048345; and WO 00/42219); and other methods or combinations of theabove methods known to those of skill in the art.

Exemplary nucleotide sequences that may be altered by geneticengineering include, but are not limited to, those categorized below.

1. Transgenes that Confer Resistance to Insects or Disease and thatEncode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones et al., Science 266: 789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262: 1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinoset al., Cell 78: 1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae), McDowell & Woffenden, (2003) Trends Biotechnol.21(4): 178-83 and Toyoda et al., (2002) Transgenic Res. 11(6):567-82. Aplant resistant to a disease is one that is more resistant to a pathogenas compared to the wild type plant.

(B) A Bacillus thuringiensis protein, a derivative thereof or asynthetic polypeptide modeled thereon. See, for example, Geiser et al.,Gene 48: 109 (1986), who disclose the cloning and nucleotide sequence ofa Bt delta-endotoxin gene. Moreover, DNA molecules encodingdelta-endotoxin genes can be purchased from American Type CultureCollection (Rockville, Md.), for example, under ATCC Accession Nos.40098, 67136, 31995 and 31998. Other non-limiting examples of Bacillusthuringiensis transgenes being genetically engineered are given in thefollowing patents and patent applications and hereby are incorporated byreference for this purpose: U.S. Pat. Nos. 5,188,960; 5,689,052;5,880,275; 5,986,177; 7,105,332; 7,208,474; WO 91/14778; WO 99/31248; WO01/12731; WO 99/24581; WO 97/40162 and U.S. application Ser. Nos.10/032,717; 10/414,637; 11/018,615; 11/404,297; 11/404,638; 11/471,878;11/780,501; 11/780,511; 11/780,503; 11/953,648; 11/953,648; and Ser. No.11/957,893.

(C) An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344: 458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

(D) An insect-specific peptide which, upon expression, disrupts thephysiology of the affected pest. For example, see the disclosures ofRegan, J. Biol. Chem. 269: 9 (1994) (expression cloning yields DNAcoding for insect diuretic hormone receptor); Pratt et al., Biochem.Biophys. Res. Comm. 163: 1243 (1989) (an allostatin is identified inDiploptera puntata); Chattopadhyay et al. (2004) Critical Reviews inMicrobiology 30 (1): 33-54 2004; Zjawiony (2004) J Nat Prod 67 (2):300-310; Carlini and Grossi-de-Sa (2002) Toxicon, 40 (11): 1515-1539;Ussuf et al. (2001) Curr Sci. 80 (7): 847-853; and Vasconcelos &Oliveira (2004) Toxicon 44 (4): 385-403. See also U.S. Pat. No.5,266,317 to Tomalski et al., who disclose genes encodinginsect-specific toxins.

(E) An enzyme responsible for a hyperaccumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

(F) An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23: 691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hookworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21: 673 (1993), who provide the nucleotide sequenceof the parsley ubi4-2 polyubiquitin gene, and U.S. Pat. Nos. 6,563,020;7,145,060 and 7,087,810.

(G) A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24: 757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104: 1467 (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

(H) A hydrophobic moment peptide. See PCT application WO 95/16776 andU.S. Pat. No. 5,580,852 disclosure of peptide derivatives of Tachyplesinwhich inhibit fungal plant pathogens) and PCT application WO 95/18855and U.S. Pat. No. 5,607,914 (teaches synthetic antimicrobial peptidesthat confer disease resistance).

(I) A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure by Jaynes et al., Plant Sci. 89: 43 (1993),of heterologous expression of a cecropin-beta lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

(J) A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. Rev. Phytopathol.28: 451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

(K) An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et al., Abstract #497, SEVENTH INT'L SYMPOSIUM ON MOLECULARPLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

(L) A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366: 469 (1993), which shows that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

(M) A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo alpha-1,4-D-polygalacturonasesfacilitate fungal colonization and plant nutrient release bysolubilizing plant cell wall homo-alpha-1,4-D-galacturonase. See Lamb etal., Bio/Technology 10: 1436 (1992). The cloning and characterization ofa gene which encodes a bean endopolygalacturonase-inhibiting protein isdescribed by Toubart et al., Plant J. 2: 367 (1992).

(N) A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10: 305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

(O) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, S., Current Biology, 5(2)(1995), Pieterse & Van Loon (2004) Curr. Opin. Plant Bio. 7(4):456-64and Somssich (2003) Cell 113(7):815-6.

(P) Antifungal genes (Cornelissen and Melchers, Pl. Physiol.101:709-712, (1993) and Parijs et al., Planta 183:258-264, (1991) andBushnell et al., Can. J. of Plant Path. 20(2):137-149 (1998). Also seeU.S. application Ser. Nos. 09/950,933; 11/619,645; 11/657,710;11/748,994; 11/774,121 and U.S. Pat. Nos. 6,891,085 and 7,306,946.

(Q) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see U.S. Pat. Nos. 5,716,820; 5,792,931; 5,798,255;5,846,812; 6,083,736; 6,538,177; 6,388,171 and 6,812,380.

(R) Cystatin and cysteine proteinase inhibitors. See U.S. Pat. No.7,205,453.

(S) Defensin genes. See WO03000863 and U.S. Pat. Nos. 6,911,577;6,855,865; 6,777,592 and 7,238,781.

(T) Genes conferring resistance to nematodes. See e.g. PCT ApplicationWO96/30517; PCT Application WO93/19181, WO 03/033651 and Urwin et al.,Planta 204:472-479 (1998), Williamson (1999) Curr Opin Plant Bio.2(4):327-31; and U.S. Pat. Nos. 6,284,948 and 7,301,069.

(U) Genes that confer resistance to Phytophthora Root Rot, such as theRps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps genes.See, for example, Shoemaker et al, Phytophthora Root Rot Resistance GeneMapping in Soybean, Plant Genome IV Conference, San Diego, Calif.(1995).

(V) Genes that confer resistance to Brown Stem Rot, such as described inU.S. Pat. No. 5,689,035 and incorporated by reference for this purpose.

(W) Genes that confer resistance to Colletotrichum, such as described inUS Patent publication US20090035765 and incorporated by reference forthis purpose. This includes the Rcg locus that may be utilized as asingle locus conversion.

2. Transgenes that Confer Tolerance to A Herbicide, for Example:

(A) A herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7: 1241 (1988), and Miki et al., Theor. Appl. Genet. 80: 449(1990), respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659;5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107;5,928,937; and 5,378,824; U.S. application Ser. No. 11/683,737, andinternational publication WO 96/33270.

(B) Glyphosate (tolerance imparted by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus phosphinothricin acetyl transferase (bar) genes), andpyridinoxy or phenoxy proprionic acids and cyclohexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah et al., which discloses the nucleotide sequence of a form of EPSPSwhich can confer glyphosate tolerance. U.S. Pat. No. 5,627,061 to Barryet al. also describes genes encoding EPSPS enzymes. See also U.S. Pat.Nos. 6,566,587; 6,338,961; 6,248,876 B1; 6,040,497; 5,804,425;5,633,435; 5,145,783; 4,971,908; 5,312,910; 5,188,642; 4,940,835;5,866,775; 6,225,114 B1; 6,130,366; 5,310,667; 4,535,060; 4,769,061;5,633,448; 5,510,471; Re. 36,449; RE 37,287 E; and U.S. Pat. No.5,491,288; and international publications EP1173580; WO 01/66704;EP1173581 and EP1173582, which are incorporated herein by reference forthis purpose. Glyphosate tolerance is also imparted to plants thatexpress a gene that encodes a glyphosate oxido-reductase enzyme asdescribed more fully in U.S. Pat. Nos. 5,776,760 and 5,463,175, whichare incorporated herein by reference for this purpose. In additionglyphosate tolerance can be imparted to plants by the over expression ofgenes encoding glyphosate N-acetyltransferase. See, for example, U.S.application Ser. Nos. 10/427,692; 10/835,615 and 11/507,751. A DNAmolecule encoding a mutant aroA gene can be obtained under ATCCaccession No. 39256, and the nucleotide sequence of the mutant gene isdisclosed in U.S. Pat. No. 4,769,061 to Comai. European PatentApplication No. 0 333 033 to Kumada et al. and U.S. Pat. No. 4,975,374to Goodman et al. disclose nucleotide sequences of glutamine synthetasegenes which confer tolerance to herbicides such as L-phosphinothricin.The nucleotide sequence of a phosphinothricin-acetyl-transferase gene isprovided in European Patent No. 0 242 246 and 0 242 236 to Leemans etal. De Greef et al., Bio/Technology 7: 61 (1989), describe theproduction of transgenic plants that express chimeric bar genes codingfor phosphinothricin acetyl transferase activity. See also, U.S. Pat.Nos. 5,969,213; 5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236;5,648,477; 5,646,024; 6,177,616 B1; and 5,879,903, which areincorporated herein by reference for this purpose. Exemplary genesconferring resistance to phenoxy proprionic acids and cyclohexones, suchas sethoxydim and haloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3 genesdescribed by Marshall et al., Theor. Appl. Genet. 83: 435 (1992).

(C) A herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3: 169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441 and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J. 285:173 (1992).

(D) Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants (see, e.g., Hattori et al. (1995)Mol Gen Genet 246:419). Other genes that confer tolerance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota et al. (1994)Plant Physiol 106:17), genes for glutathione reductase and superoxidedismutase (Aono et al. (1995) Plant Cell Physiol 36:1687, and genes forvarious phosphotransferases (Datta et al. (1992) Plant Mol Biol 20:619).

(E) Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are tolerant to theseherbicides are described in U.S. Pat. Nos. 6,288,306 B1; 6,282,837 B1;and 5,767,373; and international publication WO 01/12825.

3. Transgenes that Confer or Contribute to an Altered GrainCharacteristic, Such As:

(A) Altered fatty acids, for example, by

-   -   (1) Down-regulation of stearoyl-ACP desaturase to increase        stearic acid content of the plant. See Knultzon et al., Proc.        Natl. Acad. Sci. USA 89: 2624 (1992) and WO99/64579 (Genes for        Desaturases to Alter Lipid Profiles in Corn),    -   (2) Elevating oleic acid via FAD-2 gene modification and/or        decreasing linolenic acid via FAD-3 gene modification (see U.S.        Pat. Nos. 6,063,947; 6,323,392; 6,372,965 and WO 93/11245),    -   (3) Altering conjugated linolenic or linoleic acid content, such        as in WO 01/12800,    -   (4) Altering LEC1, AGP, Dek1, Superal1, mi1ps, various Ipa genes        such as Ipa1, Ipa3, hpt or hggt. For example, see WO 02/42424,        WO 98/22604, WO 03/011015, WO02/057439, WO03/011015, U.S. Pat.        Nos. 6,423,886, 6,197,561, 6,825,397, and U.S. Application        Serial Nos. US2003/0079247, US2003/0204870, and        Rivera-Madrid, R. et al. Proc. Natl. Acad. Sci. 92:5620-5624        (1995).

B) Altered phosphorus content, for example, by the

-   -   (1) Introduction of a phytase-encoding gene would enhance        breakdown of phytate, adding more free phosphate to the        transformed plant. For example, see Van Hartingsveldt et al.,        Gene 127: 87 (1993), for a disclosure of the nucleotide sequence        of an Aspergillus niger phytase gene.    -   (2) Modulating a gene that reduces phytate content. In maize,        this, for example, could be accomplished, by cloning and then        re-introducing DNA associated with one or more of the alleles,        such as the LPA alleles, identified in maize mutants        characterized by low levels of phytic acid, such as in WO        05/113778 and/or by altering inositol kinase activity as in WO        02/059324, US2003/0009011, WO 03/027243, US2003/0079247, WO        99/05298, U.S. Pat. No. 6,197,561, U.S. Pat. No. 6,291,224, U.S.        Pat. No. 6,391,348, WO2002/059324, US2003/0079247, Wo98/45448,        WO99/55882, WO01/04147.

(C) Altered carbohydrates affected, for example, by altering a gene foran enzyme that affects the branching pattern of starch or, a genealtering thioredoxin such as NTR and/or TRX (see. (See U.S. Pat. No.6,531,648 which is incorporated by reference for this purpose) and/or agamma zein knock out or mutant such as cs27 or TUSC27 or en27 (See U.S.Pat. No. 6,858,778 and US2005/0160488, US2005/0204418; which areincorporated by reference for this purpose). See Shiroza et al., J.Bacteriol. 170: 810 (1988) (nucleotide sequence of Streptococcus mutansfructosyltransferase gene), Steinmetz et al., Mol. Gen. Genet. 200: 220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene), Penet al., Bio/Technology 10: 292 (1992) (production of transgenic plantsthat express Bacillus licheniformis alpha-amylase), Elliot et al., PlantMolec. Biol. 21: 515 (1993) (nucleotide sequences of tomato invertasegenes), Søgaard et al., J. Biol. Chem. 268: 22480 (1993) (site-directedmutagenesis of barley alpha-amylase gene), and Fisher et al., PlantPhysiol. 102: 1045 (1993) (maize endosperm starch branching enzyme II),WO 99/10498 (improved digestibility and/or starch extraction throughmodification of UDP-D-xylose 4-epimerase, Fragile 1 and 2, Ref1, HCHL,C4H), U.S. Pat. No. 6,232,529 (method of producing high oil seed bymodification of starch levels (AGP)). The fatty acid modification genesmentioned herein may also be used to affect starch content and/orcomposition through the interrelationship of the starch and oilpathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see U.S. Pat. No. 6,787,683,US2004/0034886 and WO 00/68393 involving the manipulation of antioxidantlevels, and WO 03/082899 through alteration of a homogentisate geranylgeranyl transferase (hggt).

(E) Altered essential seed amino acids. For example, see U.S. Pat. No.6,127,600 (method of increasing accumulation of essential amino acids inseeds), U.S. Pat. No. 6,080,913 (binary methods of increasingaccumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389(high lysine), WO99/40209 (alteration of amino acid compositions inseeds), WO99/29882 (methods for altering amino acid content ofproteins), U.S. Pat. No. 5,850,016 (alteration of amino acidcompositions in seeds), WO98/20133 (proteins with enhanced levels ofessential amino acids), U.S. Pat. No. 5,885,802 (high methionine), U.S.Pat. No. 5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plantamino acid biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increasedlysine and threonine), U.S. Pat. No. 6,441,274 (plant tryptophansynthase beta subunit), U.S. Pat. No. 6,346,403 (methionine metabolicenzymes), U.S. Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414(increased methionine), WO98/56935 (plant amino acid biosyntheticenzymes), WO98/45458 (engineered seed protein having higher percentageof essential amino acids), WO98/42831 (increased lysine), U.S. Pat. No.5,633,436 (increasing sulfur amino acid content), U.S. Pat. No.5,559,223 (synthetic storage proteins with defined structure containingprogrammable levels of essential amino acids for improvement of thenutritional value of plants), WO96/01905 (increased threonine),WO95/15392 (increased lysine), US2003/0163838, US2003/0150014,US2004/0068767, U.S. Pat. No. 6,803,498, WO01/79516.

4. Genes that Control Male-Sterility:

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar et al. and chromosomal translocations asdescribed by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Inaddition to these methods, Albertsen et al., U.S. Pat. No. 5,432,068,describe a system of nuclear male sterility which includes: identifyinga gene which is needed for male fertility; silencing this native genewhich is needed for male fertility; removing the native promoter fromthe essential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing, or turning “on”,the promoter, which in turn allows the gene that confers male fertilityto be transcribed.

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN—Ac-PPT (WO 01/29237).

(B) Introduction of various stamen-specific promoters (WO 92/13956, WO92/13957).

(C) Introduction of the barnase and the barstar gene (Paul et al. PlantMol. Biol. 19:611-622, 1992).

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341; 6,297,426; 5,478,369;5,824,524; 5,850,014; and 6,265,640; all of which are herebyincorporated by reference.

5. Genes that create a site for site specific DNA integration. Thisincludes the introduction of FRT sites that may be used in the FLP/FRTsystem and/or Lox sites that may be used in the Cre/Loxp system. Forexample, see Lyznik, et al., Site-Specific Recombination for GeneticEngineering in Plants, Plant Cell Rep (2003) 21:925-932 and WO 99/25821which are hereby incorporated by reference. Other systems that may beused include the Gin recombinase of phage Mu (Maeser et al., 1991; VickiChandler, The Maize Handbook Ch. 118 (Springer-Verlag 1994), the Pinrecombinase of E. coli (Enomoto et al., 1983), and the R/RS system ofthe pSR1 plasmid (Araki et al., 1992).6. Genes that affect abiotic stress resistance (including but notlimited to flowering, ear and seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress. For example,see: WO 00/73475 where water use efficiency is altered throughalteration of malate; U.S. Pat. Nos. 5,892,009; 5,965,705; 5,929,305;5,891,859; 6,417,428; 6,664,446; 6,706,866; 6,717,034; 6,801,104;WO2000060089; WO2001026459; WO2001035725; WO2001034726; WO2001035727;WO2001036444; WO2001036597; WO2001036598; WO2002015675; WO2002017430;WO2002077185; WO2002079403; WO2003013227; WO2003013228; WO2003014327;WO2004031349; WO2004076638; WO9809521; and WO9938977 describing genes,including CBF genes and transcription factors effective in mitigatingthe negative effects of freezing, high salinity, and drought on plants,as well as conferring other positive effects on plant phenotype;US2004/0148654 and WO01/36596 where abscisic acid is altered in plantsresulting in improved plant phenotype such as increased yield and/orincreased tolerance to abiotic stress; WO2000/006341, WO04/090143, U.S.application Ser. Nos. 10/817,483 and 09/545,334 where cytokininexpression is modified resulting in plants with increased stresstolerance, such as drought tolerance, and/or increased yield. Also seeWO0202776, WO2003052063, JP2002281975, U.S. Pat. No. 6,084,153,WO0164898, U.S. Pat. No. 6,177,275, and U.S. Pat. No. 6,107,547(enhancement of nitrogen utilization and altered nitrogenresponsiveness). For ethylene alteration, see US20040128719,US20030166197 and WO200032761. For plant transcription factors ortranscriptional regulators of abiotic stress, see e.g. US20040098764 orUS20040078852.

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants, see e.g.WO97/49811 (LHY), WO98/56918 (ESD4), WO97/10339 and U.S. Pat. No.6,573,430 (TFL), U.S. Pat. No. 6,713,663 (FT), WO96/14414 (CON),WO96/38560, WO01/21822 (VRN1), WO00/44918 (VRN2), WO99/49064 (GI),WO00/46358 (FRI), WO97/29123, U.S. Pat. No. 6,794,560, U.S. Pat. No.6,307,126 (GAI), WO99/09174 (D8 and Rht), WO2004076638 and WO2004031349(transcription factors).

Using PH2FRV to Develop Another Maize Plant

Maize varieties such as PH2FRV are typically developed for use in theproduction of hybrid maize varieties. However, varieties such as PH2FRValso provide a source of breeding material that may be used to developnew maize inbred varieties. Plant breeding techniques known in the artand used in a maize plant breeding program include, but are not limitedto, recurrent selection, mass selection, bulk selection, mass selection,backcrossing, pedigree breeding, open pollination breeding, restrictionfragment length polymorphism enhanced selection, genetic marker enhancedselection, making double haploids, and transformation. Oftencombinations of these techniques are used. The development of maizehybrids in a maize plant breeding program requires, in general, thedevelopment of homozygous inbred varieties, the crossing of thesevarieties, and the evaluation of the crosses. There are many analyticalmethods available to evaluate the result of a cross. The oldest and mosttraditional method of analysis is the observation of phenotypic traitsbut genotypic analysis may also be used.

This invention is also directed to methods for producing a maize plantby crossing a first parent maize plant with a second parent maize plantwherein either the first or second parent maize plant is a maize plantof the variety PH2FRV. The other parent may be any other maize plant,such as another inbred variety or a plant that is part of a synthetic ornatural population. Any such methods using the maize variety PH2FRV arepart of this invention: selfing, sibbing, backcrosses, mass selection,pedigree breeding, bulk selection, hybrid production, crosses topopulations, and the like. These methods are well known in the art andsome of the more commonly used breeding methods are described below.Descriptions of breeding methods can also be found in one of severalreference books (e.g., Allard, Principles of Plant Breeding, 1960;Simmonds, Principles of Crop Improvement, 1979; Fehr, “Breeding Methodsfor Cultivar Development”, Production and Uses, 2^(nd) ed., Wilcoxeditor, 1987 the disclosure of which is incorporated herein byreference).

Pedigree Breeding

Pedigree breeding starts with the crossing of two genotypes, such asPH2FRV and one other inbred variety having one or more desirablecharacteristics that is lacking or which complements PH2FRV. If the twooriginal parents do not provide all the desired characteristics, othersources can be included in the breeding population. In the pedigreemethod, superior plants are selfed and selected in successive filialgenerations. In the succeeding filial generations the heterozygouscondition gives way to homogeneous varieties as a result ofself-pollination and selection. Typically in the pedigree method ofbreeding, five or more successive filial generations of selfing andselection is practiced: F1→F2; F2→F3; F3→F4; F4→F5, etc. After asufficient amount of inbreeding, successive filial generations willserve to increase seed of the developed inbred. Preferably, the inbredvariety comprises homozygous alleles at about 95% or more of its loci.

Recurrent Selection and Mass Selection

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. PH2FRV is suitable for use in arecurrent selection program. The method entails individual plants crosspollinating with each other to form progeny. The progeny are grown andthe superior progeny selected by any number of selection methods, whichinclude individual plant, half-sib progeny, full-sib progeny, selfedprogeny and topcrossing. The selected progeny are cross pollinated witheach other to form progeny for another population. This population isplanted and again superior plants are selected to cross pollinate witheach other. Recurrent selection is a cyclical process and therefore canbe repeated as many times as desired. The objective of recurrentselection is to improve the traits of a population. The improvedpopulation can then be used as a source of breeding material to obtaininbred varieties to be used in hybrids or used as parents for asynthetic cultivar. A synthetic cultivar is the resultant progeny formedby the intercrossing of several selected inbreds.

PH2FRV is suitable for use in mass selection. Mass selection is a usefultechnique when used in conjunction with molecular marker enhancedselection. In mass selection seeds from individuals are selected basedon phenotype and/or genotype. These selected seeds are then bulked andused to grow the next generation. Bulk selection requires growing apopulation of plants in a bulk plot, allowing the plants toself-pollinate, harvesting the seed in bulk and then using a sample ofthe seed harvested in bulk to plant the next generation. Instead of selfpollination, directed pollination could be used as part of the breedingprogram.

Mutation Breeding

Mutation breeding is one of many methods that could be used to introducenew traits into PH2FRV. PH2FRV is suitable for use in a mutationbreeding program. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation; such as X-rays, Gamma rays (e.g. cobalt60 or cesium 137), neutrons, (product of nuclear fission by uranium 235in an atomic reactor), Beta radiation (emitted from radioisotopes suchas phosphorus 32 or carbon 14), or ultraviolet radiation (preferablyfrom 2500 to 2900 nm), or chemical mutagens (such as base analogues(5-bromo-uracil), related compounds (8-ethoxy caffeine), antibiotics(streptonigrin), alkylating agents (sulfur mustards, nitrogen mustards,epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones),azide, hydroxylamine, nitrous acid, or acridines. Once a desired traitis observed through mutagenesis the trait may then be incorporated intoexisting germplasm by traditional breeding techniques, such asbackcrossing. Details of mutation breeding can be found in “Principlesof Cultivar Development” Fehr, 1993 Macmillan Publishing Company, thedisclosure of which is incorporated herein by reference. In addition,mutations created in other varieties may be used to produce a backcrossconversion of PH2FRV that comprises such mutation.

Production of Double Haploids

The production of double haploids can also be used for the developmentof inbreds in the breeding program. For example, an F1 hybrid for whichPH2FRV is a parent can be used to produce double haploid plants. Doublehaploids are produced by the doubling of a set of chromosomes (1N) froma heterozygous plant to produce a completely homozygous individual. Forexample, see Wan et al., “Efficient Production of Doubled Haploid PlantsThrough Colchicine Treatment of Anther-Derived Maize Callus”,Theoretical and Applied Genetics, 77:889-892, 1989 and US2003/0005479.This can be advantageous because the process omits the generations ofselfing needed to obtain a homozygous plant from a heterozygous source.

Haploid induction systems have been developed for various plants toproduce haploid tissues, plants and seeds. The haploid induction systemcan produce haploid plants from any genotype by crossing a selectedvariety (as female) with an inducer variety. Such inducer varieties formaize include Stock 6 (Coe, 1959, Am. Nat. 93:381-382; Sharkar and Coe,1966, Genetics 54:453-464) RWS (available online from the UniversitätHohenheim), KEMS (Deimling, Roeber, and Geiger, 1997, Vortr.Pflanzenzuchtg 38:203-224), KMS and ZMS (Chalyk, Bylich & Chebotar,1994, MNL 68:47; Chalyk & Chebotar, 2000, Plant Breeding 119:363-364),and indeterminate gametophyte (ig) mutation (Kermicle 1969 Science166:1422-1424). The disclosures of which are incorporated herein byreference.

Methods for obtaining haploid plants are also disclosed in Kobayashi, M.et al., Journ. of Heredity 71(1):9-14, 1980, Pollacsek, M., Agronomie(Paris) 12(3):247-251, 1992; Cho-Un-Haing et al., Journ. of Plant Biol.,1996, 39(3):185-188; Verdoodt, L., et al., February 1998, 96(2):294-300;Genetic Manipulation in Plant Breeding, Proceedings InternationalSymposium Organized by EUCARPIA, Sep. 8-13, 1985, Berlin, Germany;Chalyk et al., 1994, Maize Genet Coop. Newsletter 68:47; Chalyk, S. T.,1999, Maize Genet. Coop. Newsletter 73:53-54; Coe, R. H., 1959, Am. Nat.93:381-382; Deimling, S. et al., 1997, Vortr. Pflanzenzuchtg 38:203-204;Kato, A., 1999, J. Hered. 90:276-280; Lashermes, P. et al., 1988, Theor.Appl. Genet. 76:570-572 and 76:405-410; Tyrnov, V. S. et al., 1984,Dokl. Akad. Nauk. SSSR 276:735-738; Zabirova, E. R. et al., 1996,Kukuruza I Sorgo N4, 17-19; Aman, M. A., 1978, Indian J. Genet PlantBreed 38:452-457; Chalyk S. T., 1994, Euphytica 79:13-18; Chase, S. S.,1952, Agron. J. 44:263-267; Coe, E. H., 1959, Am. Nat. 93:381-382; Coe,E. H., and Sarkar, K. R., 1964 J. Hered. 55:231-233; Greenblatt, I. M.and Bock, M., 1967, J. Hered. 58:9-13; Kato, A., 1990, Maize Genet.Coop. Newsletter 65:109-110; Kato, A., 1997, Sex. Plant Reprod.10:96-100; Nanda, D. K. and Chase, S. S., 1966, Crop Sci. 6:213-215;Sarkar, K. R. and Coe, E. H., 1966, Genetics 54:453-464; Sarkar, K. R.and Coe, E. H., 1971, Crop Sci. 11:543-544; Sarkar, K. R. and Sachan J.K. S., 1972, Indian J. Agric. Sci. 42:781-786; Kermicle J. L., 1969,Mehta Yeshwant, M. R., Genetics and Molecular Biology, September 2000,23(3):617-622; Tahir, M. S. et al. Pakistan Journal of Scientific andIndustrial Research, August 2000, 43(4):258-261; Knox, R. E. et al.Plant Breeding, August 2000, 119(4):289-298; U.S. Pat. No. 5,639,951 andU.S. patent application Ser. No. 10/121,200, the disclosures of whichare incorporated herein by reference.

Thus, an embodiment of this invention is a process for making ahomozygous PH2FRV progeny plant substantially similar to PH2FRV byproducing or obtaining a seed from the cross of PH2FRV and another maizeplant and applying double haploid methods to the F1 seed or F1 plant orto any successive filial generation. Such methods decrease the number ofgenerations required to produce an inbred with similar genetics orcharacteristics to PH2FRV. See Bernardo, R. and Kahler, A. L., Theor.Appl. Genet. 102:986-992, 2001.

In particular, a process of making seed substantially retaining themolecular marker profile of maize variety PH2FRV is contemplated, suchprocess comprising obtaining or producing F1 hybrid seed for which maizevariety PH2FRV is a parent, inducing double haploids to create progenywithout the occurrence of meiotic segregation, obtaining the molecularmarker profile of maize variety PH2FRV, and selecting progeny thatretain the molecular marker profile of PH2FRV.

Another embodiment of the invention is a maize seed derived from inbredmaize variety PH2FRV produced by crossing a plant or plant part ofinbred maize variety PH2FRV with another plant, wherein representativeseed of said inbred maize variety PH2FRV has been deposited and whereinsaid maize seed derived from the inbred maize variety PH2FRV has 92%,93%, 94%, 95%, 96%, 97%, 98% or 99% of the same polymorphisms formolecular markers as the plant or plant part of inbred maize varietyPH2FRV. Sequences for the public markers can be found, for example, inthe Panzea database which is available online from Panzea. The type ofmolecular marker used in the molecular profile can be but is not limitedto Single Nucleotide Polymorphisms, SNPs. A maize seed derived frominbred maize variety PH2FRV produced by crossing a plant or plant partof inbred maize variety PH2FRV with another plant, whereinrepresentative seed of said inbred maize variety PH2FRV has beendeposited and wherein said maize seed derived from the inbred maizevariety PH2FRV has essentially the same morphological characteristics asmaize variety PH2FRV when grown in the same environmental conditions.The same environmental conditions may be, but is not limited to aside-by-side comparison. The characteristics can be those listed inTable 1. The comparison can be made using any number of professionallyaccepted experimental designs and statistical analysis.

Use of PH2FRV in Tissue Culture

This invention is also directed to the use of PH2FRV in tissue culture.As used herein, the term “tissue culture” includes plant protoplasts,plant cell tissue culture, cultured microspores, plant calli, plantclumps, and the like. As used herein, phrases such as “growing the seed”or “grown from the seed” include embryo rescue, isolation of cells fromseed for use in tissue culture, as well as traditional growing methods.

Duncan, Williams, Zehr, and Widholm, Planta (1985) 165:322-332 reflectsthat 97% of the plants cultured that produced callus were capable ofplant regeneration. Subsequent experiments with both inbreds and hybridsproduced 91% regenerable callus that produced plants. In a further studyin 1988, Songstad, Duncan & Widholm in Plant Cell Reports (1988),7:262-265 reports several media additions that enhance regenerability ofcallus of two inbred varieties. Other published reports also indicatedthat “nontraditional” tissues are capable of producing somaticembryogenesis and plant regeneration. K. P. Rao, et al., Maize GeneticsCooperation Newsletter, 60:64-65 (1986), refers to somatic embryogenesisfrom glume callus cultures and B. V. Conger, et al., Plant Cell Reports,6:345-347 (1987) indicates somatic embryogenesis from the tissuecultures of maize leaf segments. Thus, it is clear from the literaturethat the state of the art is such that these methods of obtaining plantsare, and were, “conventional” in the sense that they are routinely usedand have a very high rate of success.

Tissue culture of maize, including tassel/anther culture, is describedin U.S. 2002/0062506A1 and European Patent Application, publicationEP0160,390, each of which are incorporated herein by reference for thispurpose. Maize tissue culture procedures are also described in Green andRhodes, “Plant Regeneration in Tissue Culture of Maize,” Maize forBiological Research (Plant Molecular Biology Association,Charlottesville, Va. 1982, at 367-372) and in Duncan, et al., “TheProduction of Callus Capable of Plant Regeneration from Immature Embryosof Numerous Zea Mays Genotypes,” 165 Planta 322-332 (1985). Thus,another aspect of this invention is to provide cells which upon growthand differentiation produce maize plants having the genotype and/orphenotypic characteristics of variety PH2FRV.

Seed Treatments and Cleaning

Another embodiment of this invention is the method of harvesting theseed of the maize variety PH2FRV as seed for planting. Embodimentsinclude cleaning the seed, treating the seed, and/or conditioning theseed. Cleaning the seed includes removing foreign debris such as weedseed and removing chaff, plant matter, from the seed. Conditioning theseed can include controlling the temperature and rate of dry down andstoring seed in a controlled temperature environment. Seed treatment isthe application of a composition to the seed such as a coating orpowder. Methods for producing a treated seed include the step ofapplying a composition to the seed or seed surface. Some examples ofcompositions are insecticides, fungicides, pesticides, antimicrobials,germination inhibitors, germination promoters, cytokinins, andnutrients.

To protect and to enhance yield production and trait technologies, seedtreatment options can provide additional crop plan flexibility and costeffective control against insects, weeds and diseases, thereby furtherenhancing the invention described herein. Seed material can be treated,typically surface treated, with a composition comprising combinations ofchemical or biological herbicides, herbicide safeners, insecticides,fungicides, germination inhibitors and enhancers, nutrients, plantgrowth regulators and activators, bactericides, nematicides, avicidesand/or molluscicides. These compounds are typically formulated togetherwith further carriers, surfactants or application-promoting adjuvantscustomarily employed in the art of formulation. The coatings may beapplied by impregnating propagation material with a liquid formulationor by coating with a combined wet or dry formulation. Examples of thevarious types of compounds that may be used as seed treatments areprovided in The Pesticide Manual: A World Compendium, C.D.S. Tomlin Ed.,Published by the British Crop Production Council, which is herebyincorporated by reference.

Some seed treatments that may be used on crop seed include, but are notlimited to, one or more of abscisic acid, acibenzolar-S-methyl,avermectin, amitrol, azaconazole, azospirillum, azadirachtin,azoxystrobin, Bacillus spp. (including one or more of cereus, firmus,megaterium, pumilis, sphaericus, subtilis and/or thuringiensis),Bradyrhizobium spp. (including one or more of betae, canariense,elkanii, iriomotense, japonicum, liaonigense, pachyrhizi and/oryuanmingense), captan, carboxin, chitosan, clothianidin, copper,cyazypyr, difenoconazole, etidiazole, fipronil, fludioxonil,fluoxastrobin, fluquinconazole, flurazole, fluxofenim, harpin protein,imazalil, imidacloprid, ipconazole, isoflavenoids,lipo-chitooligosaccharide, mancozeb, manganese, maneb, mefenoxam,metalaxyl, metconazole, myclobutanil, PCNB, penflufen, penicillium,penthiopyrad, permethrine, picoxystrobin, prothioconazole,pyraclostrobin, rynaxypyr, S-metolachlor, saponin, sedaxane, TCMTB,tebuconazole, thiabendazole, thiamethoxam, thiocarb, thiram,tolclofos-methyl, triadimenol, trichoderma, trifloxystrobin,triticonazole and/or zinc. PCNB seed coat refers to EPA registrationnumber 00293500419, containing quintozen and terrazole. TCMTB refers to2-(thiocyanomethylthio) benzothiazole.

Seed varieties and seeds with specific transgenic traits may be testedto determine which seed treatment options and application rates maycomplement such varieties and transgenic traits in order to enhanceyield. For example, a variety with good yield potential but head smutsusceptibility may benefit from the use of a seed treatment thatprovides protection against head smut, a variety with good yieldpotential but cyst nematode susceptibility may benefit from the use of aseed treatment that provides protection against cyst nematode, and soon. Likewise, a variety encompassing a transgenic trait conferringinsect resistance may benefit from the second mode of action conferredby the seed treatment, a variety encompassing a transgenic traitconferring herbicide resistance may benefit from a seed treatment with asafener that enhances the plants resistance to that herbicide, etc.Further, the good root establishment and early emergence that resultsfrom the proper use of a seed treatment may result in more efficientnitrogen use, a better ability to withstand drought and an overallincrease in yield potential of a variety or varieties containing acertain trait when combined with a seed treatment.

INDUSTRIAL APPLICABILITY

Another embodiment, is a method of harvesting the grain of the F1 plantof variety PH2FRV and using the grain in a commodity. Methods ofproducing a commodity plant product are also provided. Examples of maizegrain as a commodity plant product include, but are not limited to,oils, meals, flour, starches, syrups, proteins, cellulose, silage, andsugars. Maize grain is used as human food, livestock feed, and as rawmaterial in industry. The food uses of maize, in addition to humanconsumption of maize kernels, include both products of dry- andwet-milling industries. The principal products of maize dry milling aregrits, meal and flour. The maize wet-milling industry can provide maizestarch, maize syrups, and dextrose for food use. Maize oil is recoveredfrom maize germ, which is a by-product of both dry- and wet-millingindustries. Processing the grain can include one or more of cleaning toremove foreign material and debris from the grain, conditioning, such asaddition of moisture to the grain, steeping the grain, wet milling, drymilling and sifting.

Maize, including both grain and non-grain portions of the plant, is alsoused extensively as livestock feed, primarily for beef cattle, dairycattle, hogs, and poultry.

Industrial uses of maize include production of ethanol, maize starch inthe wet-milling industry and maize flour in the dry-milling industry.The industrial applications of maize starch and flour are based onfunctional properties, such as viscosity, film formation, adhesiveproperties, and ability to suspend particles. The maize starch and flourhave application in the paper and textile industries. Other industrialuses include applications in adhesives, building materials, foundrybinders, laundry starches, explosives, oil-well muds, and other miningapplications.

Plant parts other than the grain of maize are also used in industry: forexample, stalks and husks are made into paper and wallboard and cobs areused for fuel and to make charcoal.

The seed of maize variety PH2FRV, the plant produced from the seed, thehybrid maize plant produced from the crossing of the variety, hybridseed, and various parts of the hybrid maize plant and transgenicversions of the foregoing, can be utilized for human food, livestockfeed, and as a raw material in industry.

All publications, patents, and patent applications mentioned in thespecification are incorporated by reference herein for the purpose citedto the same extent as if each was specifically and individuallyindicated to be incorporated by reference herein.

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding. Asis readily apparent to one skilled in the art, the foregoing are onlysome of the methods and compositions that illustrate the embodiments ofthe foregoing invention. It will be apparent to those of ordinary skillin the art that variations, changes, modifications, and alterations maybe applied to the compositions and/or methods described herein withoutdeparting from the true spirit, concept, and scope of the invention.

As used herein, the terms “comprises,” “comprising,” “includes,”“including,” “has,” “having,” “contains”, “containing,” “characterizedby” or any other variation thereof, are intended to cover anon-exclusive inclusion.

Unless expressly stated to the contrary, “or” is used as an inclusiveterm. For example, a condition A or B is satisfied by any one of thefollowing: A is true (or present) and B is false (or not present), A isfalse (or not present) and B is true (or present), and both A and B aretrue (or present). The indefinite articles “a” and “an” preceding anelement or component are nonrestrictive regarding the number ofinstances (i.e., occurrences) of the element or component. Therefore “a”or “an” should be read to include one or at least one, and the singularword form of the element or component also includes the plural unlessthe number is obviously meant to be singular.

DEPOSITS

Applicant has made a deposit of at least 2,500 seeds of Maize VarietyPH2FRV with the American Type Culture Collection (ATCC), 10801University Boulevard, Manassas, Va. 20110-2209, USA, with ATCC DepositNo. PTA-123870. The seeds deposited with the ATCC on Feb. 28, 2017 wereobtained from the seed of the variety maintained by Pioneer Hi-BredInternational, Inc., 7250 NW 62^(nd) Avenue, Johnston, Iowa, 50131 sinceprior to the filing date of this application. Access to this seed willbe available during the pendency of the application to the Commissionerof Patents and Trademarks and persons determined by the Commissioner tobe entitled thereto upon request. Upon allowance of any claims in theapplication, the Applicant will make the deposit available to the publicpursuant to 37 C.F.R. §1.808. This deposit of the Maize Variety PH2FRVwill be maintained in the ATCC depository, which is a public depository,for a period of 30 years, or 5 years after the most recent request, orfor the enforceable life of the patent, whichever is longer, and will bereplaced if it becomes nonviable during that period. Additionally,Applicant has or will satisfy all of the requirements of 37 C.F.R.§§1.801-1.809, including providing an indication of the viability of thesample upon deposit. Applicant has no authority to waive anyrestrictions imposed by law on the transfer of biological material orits transportation in commerce. Applicant does not waive anyinfringement of rights granted under this patent or under the PlantVariety Protection Act (7 USC 2321 et seq.).

TABLE 1 Variety Description Information Current Variety Name PH2FRVPlant height with tassel (cm) 206 Ear height (cm) 54 Leaf width of blade(cm) 8 Leaf limb anthocyanin color Absent to Very Weak intensity ofentire plant Nodes anthocyanin color intensity absent or very weakInternode anthocyanin color intensity medium Length of main axis above17 highest lateral branch (cm) Length of main axis above lowest 26lateral branch (cm) Degree of stem zig-zag slight Ear husk length mediumEar shank length short Leaf length score 0.90 m to 1 m Leaf anglebetween blade and stem small (6 to 37 degrees) Leaf tip shape pointed torounded Foliage intensity of green color medium Leaf blade undulation ofmargin intermediate Silk anthocyanin color intensity strong Anther coloryellow Anther anthocyanin color intensity weak Glume color pink Glumeanthocyanin coloration at base medium Glume anthocyanin color mediumexcluding base Brace roots anthocyanin coloration medium to strongSheath anthocyanin color intensity weak (whole plant, ear insertionlevel) Sheath anthocyanin color intensity strong at first leaf stageLeaf sheath hairiness scale 1 (1-none, 6-fuzzy) Tassel angle betweenmain axis small and lateral branches Tassel lateral branch curvatureabsent or very slightly recurved (<5 degrees) Number of primary lateralfew (4 to 10) tassel branches Primary tassel branch length shortSecondary tassel branches (number) 0 to 3 branches Tassel spikeletdensity medium Grain Texture (Flint/Dent) flint-like Ear shape (taper)conico-cylindrical Ear length (cm) 148 Ear diameter (cm) 36 Number ofrows of grain on ear 12 Dorsal side of grain color orange Top of graincolor orange Kernel shape cuneiform Cob diameter (mm) 21 Cob color redCob anthocyanin Color Intensity medium Cob glume anthocyanin colormedium intensity

TABLE 2 Inbred PH2FRV platform BLUP breeding values Weighted Trait BLUPvalue BRLPNE 71.4 BRLPNL 82.1 EARHT 45.3 ERTLPN 89.3 GDUSHD 105.7 GDUSLK105.3 GIBERS 6.5 HDSMT 97.1 HSKCVR 6.6 LRTLPN 93.0 MST 30.1 NLFBLT 5.4PLTHT 107.1 STAGRN 5.7 STLPCN 96.1 YIELD 163.5

TABLE 3 Inbred PH2FRV as parent in hybrid BRLPNE BRLPNL EARHT ftnoteBLUP SE BLUP SE BLUP SE Hybrid1 50.6 7.5 75.8 6.5 48.3 0.6 ERTLPN GDUSHDGDUSLK ftnote BLUP SE BLUP SE BLUP SE Hybrid1 82.1 5.5 107.0 0.5 107.70.5 GIBERS HDSMT HSKCVR ftnote BLUP SE BLUP SE BLUP SE Hybrid1 6.0 0.496.9 0.6 6.7 0.6 LRTLPN MST NLFBLT ftnote BLUP SE BLUP SE BLUP SEHybrid1 95.4 4.4 31.0 0.2 5.1 0.4 PLTHT STAGRN STLPCN ftnote BLUP SEBLUP SE BLUP SE Hybrid1 104.6 0.7 6.0 0.3 96.5 4.9 YIELD ftnote BLUP SEHybrid1 169.5 1.6 a wherein inbred comprises a trait conversionconferring insect control b wherein inbred comprises a trait conversionconferring herbicide tolerance c wherein inbred comprises a traitconversion conferring disease control

What is claimed is:
 1. A seed, plant, plant part, or plant cell ofinbred maize variety PH2FRV, representative seed of the variety havingbeen deposited under ATCC accession number PTA-123870.
 2. The plant partof claim 1, wherein the plant part is an ovule or pollen.
 3. An F1hybrid maize seed of PH2FRV produced by crossing the plant or plant partof claim 1 with a different maize plant.
 4. The F1 hybrid seed of claim3, wherein the plant of maize variety PH2FRV further comprises atransgene that is inherited by the seed, wherein the transgene wasintroduced into inbred maize variety PH2FRV by backcrossing or genetictransformation.
 5. An F1 hybrid maize plant or plant part produced bygrowing the maize seed of claim
 3. 6. A method for producing a secondmaize plant, the method comprising applying plant breeding techniques tothe F1 plant or plant part of claim 5 to produce the second maize plant.7. A method for producing a second maize plant or plant part, the methodcomprising doubling haploid seed generated from a cross of the plant orplant part of claim 5 with an inducer variety, thereby producing thesecond maize plant or plant part.
 8. A method of making a commodityplant product comprising silage, starch, fat, syrup or protein, themethod comprising producing the commodity plant product from the maizeplant or plant part of claim
 5. 9. A method of producing a maize plantderived from the variety PH2FRV, comprising: a) crossing the plant ofclaim 1 with itself or a second plant to produce progeny seed; b)growing the progeny seed to produce a progeny plant and crossing theprogeny plant with itself or a different plant to produce furtherprogeny seed; and c) repeating step (b) for at least one additionalgeneration to produce a maize plant derived from the variety PH2FRV. 10.A method for producing nucleic acids, the method comprising isolatingnucleic acids from the seed, plant, plant part, or plant cell ofclaim
 1. 11. A converted seed, plant, plant part or plant cell of inbredmaize variety PH2FRV, representative seed of the maize variety PH2FRVhaving been deposited under ATCC accession number PTA-123870, whereinthe converted seed, plant, plant part or plant cell comprises a locusconversion, and wherein the plant or a plant grown from the convertedseed, plant part or plant cell comprises the locus conversion andotherwise comprises all the phenotypic characteristics of maize varietyPH2FRV when grown under the same environmental conditions.
 12. Theconverted seed, plant, plant part or plant cell of claim 11, wherein thelocus conversion confers a property selected from the group consistingof male sterility, site-specific recombination, abiotic stresstolerance, altered phosphorus, altered antioxidants, altered fattyacids, altered essential amino acids, altered carbohydrates, herbicidetolerance, insect resistance and disease resistance.
 13. A maize seedproduced by crossing the plant or plant part of claim 11 with adifferent maize plant.
 14. A maize plant or plant part produced bygrowing the seed of claim
 13. 15. A method for producing a second maizeplant, the method comprising applying plant breeding techniques to theplant or plant part of claim 14 to produce the second maize plant.
 16. Amethod for producing a second maize plant or plant part, the methodcomprising doubling haploid seed generated from a cross of the plant orplant part of claim 14 with an inducer variety, thereby producing thesecond maize plant or plant part.
 17. A method of making a commodityplant product comprising silage, starch, fat, syrup or protein, themethod comprising producing the commodity plant product from the maizeplant or plant part of claim
 14. 18. A method of producing a maize plantderived from the variety PH2FRV, comprising: a) crossing the plant ofclaim 11 with itself or a second plant to produce progeny seed; b)growing the progeny seed to produce a progeny plant and crossing theprogeny plant with itself or a different plant to produce furtherprogeny seed; and c) repeating steps (a) and (b) with sufficientinbreeding until a seed of an inbred maize plant derived from thevariety PH2FRV is produced.
 19. The method of claim 18, furthercomprising crossing the inbred maize plant derived from the varietyPH2FRV with a maize plant of a different genotype to produce seed of ahybrid plant derived from the maize variety PH2FRV.
 20. A method forproducing nucleic acids, the method comprising isolating nucleic acidsfrom the seed, plant, plant part, or plant cell of claim 11.